1. Introduction
A current challenge in the field of tissue engineering is soft tissue replacement. Techniques for soft tissue reconstruction include use of autologous fat transplantation, alloplastic implants and autologous tissue flaps. However, these approaches have disadvantages, including donor-site morbidity, implant migration, foreign body reaction and immune system rejection. The use of autologous stem cells expanded
The presence of self-renewing cells within the bone marrow of mice was reported in 1963 which was later known as hematopoietic stem cells (HSCs) (Becker, et al.;1963, Zhang, et al.;1999). Several years later, HSCs were identified in umbilical cord blood by other investigators (Aust, et al.; 2004, Dellavalle, et al.;2007). Furthermore, several other adult stem cell types such as neural stem cells (Dellavalle, et al.; 2007, Guilak, et al.;2010), were isolated and identified. Moreover, a population of plastic adherent cells were isolated from collagenase digests of adipose tissue.
Adipose tissue derived stem cells were termed as: adipose derived stem/stromal cells (ASCs), adipose-derived adult stem (ADAS) cells, adipose-derived adult stromal cells, adipose-derived stromal cells (ADSCs), adipose stromal cells (ASCs), adipose mesenchymal stem cells (AdMSCs), lipoblast, pericyte, preadipocyte, and processed lipoaspirate (PLA) cells. To prevent the confusion in the literature, the International Fat Applied Technology Society reached a consensus to adopt the term “adipose-derived stem cells” (ASCs) to identify the isolated, plastic-adherent, multipotent cell population (Gimble, et al.;2007). Adipose tissue derives from the mesodermal layer of the embryo and develops both pre- and postnatally (Gonda, et al.;2008, Kakudo, et al.;2008). Researchers studying the development of adipose tissue have long worked with a fibroblastic cell line, known as preadipocytes, isolated from adipose tissue (Cowherd, et al.;1999, Gregoire, et al.;1998, Kirkland and Hollenberg;1998, Nam and Lobie;2000, Sorisky;1999).
The reason why adipose tissue would contain a stem cell population is not still clear. There is some discussion whether the cells are subpopulation of fibroblasts reside within the fat tissue or are perhaps mesenchymal or peripheral blood stem cells passing through the fat tissue (Crisan, et al.; 2008, Dellavalle, et al.; 2007).
The ASCs represent a readily available source for isolation of potentially useful stem cells (Sterodimas, et al.; 2010). In culture, they have shown to have an impressive developmental plasticity, including the ability to undergo multilineage differentiation and self-renewal (Liu, et al.; 2009). When ASCs are compared with BM-MSCs, further similarities have been demonstrated in regards to their growth kinetics, cell senescence, gene transduction efficiency (De Ugarte, et al.; 2003), as well as CD surface marker expression (Gronthos, et al.; 2001, Katz, et al.; 2005, Zuk, et al.; 2002) and gene transcription (Katz, et al.; 2005). Compared to bone marrow MSCs, ASCs have potential advantages for tissue engineering application, because of the tissue accessibility, multipotency and ease of isolation without painful procedures or donor site injury.
In this chapter we will discuss the potential use of adipose-derived stem cells in the field of tissue engineering.
2. Isolation and expansion of ASCs
The initial methods to isolate cells from adipose tissue were pioneered by Rodbell and colleagues in the 1960s (Rodbell; 1966, Rodbell; 1966, Rodbell and Jones; 1966) using rat fat tissue. These methods were further adapted for human tissues by several other groups (Deslex, et al.; 1987, Engfeldt, et al.; 1980, Ho, et al.; 2010). The current methods for isolating ASCs rely on a collagenase digestion followed by centrifugal separation to isolate the stromal/vascular cells from primary adipocytes. The pellet is resuspended with a basal medium containing 10% foetal bovine serum (Estes, et al.; 2008). The cell suspension is filtered through 100 µm cell strainer and the cells are plated and incubated at 37ºC in the presence of 5% CO2. The medium is changed every second day until the cells reach 80-90% confluence. A large number of ASCs can be harvested in this manner, with yields of approximately 250,000 cells per gram of tissue (Aust, et al.; 2004, Guilak, et al.; 2010). In order to remove the use of animal products in human ASC cultures, a very low human serum expansion medium and a completely serum-free medium have been recently reported (Parker, et al.; 2007). Furthermore it was reported that use of platelet- rich plasma can enhance the proliferation of human ASCs. These results can support the clinical application of platelet-rich plasma for cell based, soft-tissue engineering and wound healing (Kakudo, et al.; 2008).
ASCs should be harvested at 80% confluence for freezing. Cryopreservation medium consists of 80% fetal bovine serum, 10% dimethylsulfoxide (DMSO) and 10% DMEM/Ham’s F-12. The cells should be stored in a final concentration of 1–2 million viable cells per milliliter of cryopreservation medium. Aliqouted vials are first frozen in an alcohol freezing container and store and are stored at -80ºC overnight. On the next day, the frozen vials can be transferred to a liquid nitrogen container for long-term storage. Successful storage of ASCs more than 6 months has been shown. This ensures the availability of autologous banked ASCs for clinical applications in the future (De Rosa, et al.;2009, Gonda, et al.;2008).
2.1. Characterization of ASCs
In order to characterize the undifferentiated animal or human ASC cells cultured in vitro flow cytometric and immunohistochemical methods are widely used (Gronthos, et al.; 2001, Zuk, et al.; 2001). The cell surface phenotype of ASC is quite similar to MSCs (mesenchymal stem cells). Both ASC and MSC cells express CD29, CD44, CD71, CD90, and CD105 (Zuk, et al.; 2002). In contrast, no expression of the hematopoietic lineage markers CD31, CD34 and CD45 was observed in either of the cultures. In addition the ASC cells express the neutral endopeptidase (CD10 or common acute lymphocytic leukemia antigen CALLA), aminopeptidase (CD13), and ecto nucleotidase (CD73). Furthermore, ASC cells produce Type I and Type III collagens, osteopontin, ostenectin, Thy-1 (CD90), and MUC-18 (CD146) (Gimble and Guilak; 2003).
Different investigator have reported different pattern of expression. For example, while Gronthos et al. (Gronthos, et al.;2001) detected CD34 and VCAM (CD106) on ASC cells, Zuk et al. (Zuk, et al.;2002) did not. Likewise, while Zuk et al. (Zuk, et al.;2002) detected Stro-1, Gronthos et al. did not. These discrepancies could be due to the differences in cell isolation methods, how long the cells were cultured prior to analysis and sensitivity differences between immunohistochemical and flow cytometric detection methods (Zuk, et al.;2002, Zuk, et al.;2001).
3. ASCs applications
Adipose tissue has proven to serve as an abundant source of adult stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. ASCs can be differentiated into variety of cell types. Differentiation is commonly induced by insulin, dexamethasone, cyclic AMP agonist, β-glycerophosphate, heparin, ascorbate and different cytokines depending on the lineage type.
ASCs like BM-MSCs, differentiate
3.1. ASCs differentiation
3.1.1. Adipogenesis
ASCs in response to inductive compounds including glucocorticoid receptor ligands (dexamethasone), insulin, cyclic AMP agonist (forskolin) and peroxisome proliferator-activated receptor gamma (PPARγ) undergo adipogenic differentiation (Farmer; 2006, Hauner, et al.; 1989, Lazar; 2005, Zuk, et al.; 2001). During the differentiation process ASCs reduce their proliferation rate and undergo morphological changes. ASCs are induced in the adipocyte differentiation medium containing biotin, d-pantothenate, dexamethasone, methylisobutylxanthine, insulin and equivalent PPARγ agonist. After induction for 2 weeks in adipogenic medium the human ASC contain vacuoles filled with neutral lipid cells which can be further stained for intracellular lipid droplets accumulation using an Oil Red O stain (Preece;1972).
In addition, these cells secrete increased amounts of the adipocyte protein leptin, and transcribe adipogenic mRNAs such as the fatty acid binding protein, aP2 and lipoprotein lipase (Halvorsen, et al.; 2001, Hauner, et al.; 1989, Sen, et al.; 2001). Some of these parameters such as leptin, aP2 mRNA levels were quantified and found to be increased by several hundred-fold during the differentiation process (Halvorsen, et al.; 2001, Sen, et al.; 2001). It is reported that ASCs harvested from female mice differentiate more efficiently into adipocytes than those from male mice (Ogawa, et al.; 2004).
One of the most important uses of ASCs is for the replacement of adipose tissue itself. Large soft tissue defects are a common problem following trauma, burns and oncological resections. Several studies demonstrated the
3.1.2. Smooth muscle
ASCs can be differentiated to smooth muscle cells (SMCs) and might offer a cell source for hollow organ engineering. For myogenic differentiation ASCs at passages 3 through 5 are cultured in smooth muscle inductive medium consisting of MCDB131 supplemented with 1% FBS and 100u/ml of heparin for up to 6 weeks at 37ºC with 5% CO2. The media is changed every 3 days and cell splitting is not required (Jack, et al.; 2009).
The cellular changes after differentiation can be investigated by real- time PCR at mRNA level. As reported the expression of muscle actin (SMA), calponin and myosin heavy chain showed an increase after growth in differentiation medium (Jack, et al.; 2009). The same was observed at protein levels, induction media induced differentiation of the ASCs into a smooth muscle phenotype in which the expression of smooth muscle specific proteins SMA, caldesmon, and myosin heavy chain (MHC) was increased (Jack, et al.; 2009).
Differentiation is a complex process and has a dramatical effect on cell size, shape, membrane potential, metabolic activity and responsiveness to external signals. One of the main characteristic of SMCs is their contractility which plays important roles in angiogenesis, blood vessel maintenance, and mechanical regulation of hollow organs such as bladder.
Differentiated SMCs can show two specific phenotypes, which vary from synthetic and proliferative to contractile and quiescent (Beamish, et al.; 2010). SMCs exhibit a contractile phenotype characterized by high expression of specific contractile proteins including SMA, calponin, SM22, smoothelin,
Since differentiated human ASCs express smooth muscle specific proteins they may prove to be of value in the repair of smooth muscle defects in the gastrointestinal and urinary tracts. Juan et al. reported that the ASCs from different sites show different myogenic differentiation abilities
3.1.3. Osteogenesis. Bone defect repair
In the past decade, several groups isolated cells from the adipose tissue of humans and other species capable of differentiating into osteoblasts
For osteogenic differentiation confluent ASCs cells are incubated for 3 weeks in DMEM containing 10% FBS, 100 nM dexamethasone, 10 mM β-glycerophosphate and 50 μM L-ascorbic acid-2-phosphate. After fixation cells are incubated at 37ºC for 1 hour with 0.16% naphthol AS-TR phosphate and 0.8% Fast Blue BB dissolved in 0.1 M tris buffer (pH 9.0). For osteogenic differentiation cells were also incubated in 1% alizarin red S for 3 minutes to detect calcium deposition (Sakuma, et al.; 2009).
Over a 2- 4 week period
3.1.4. Myogenesis: skeletal muscle repair
There is several line of evidence that ASC cells can differentiate along each of the myocyte lineage pathways when cultured in myogenic induction medium containing 0.1
Skeletal myogenesis is characterized by a period of myoblast proliferation, followed by the expression of muscle-specific proteins and fusion to form multinucleated myotubules. Early myogenic differentiation is characterized by the expression of several myogenic regulatory factors including myogenic determination factor MyoD1 (Weintraub, et al.; 1991). Terminally differentiated myoblasts can be characterized by the expression of myosin and the presence of multiple nuclei (Silberstein, et al.; 1986).
In the first
3.1.5. Chondrogenesis
ASCs display chondrogenic characteristics following induction with ascorbate, dexamethasone and transforming growth factor-β (Awad, et al.; 2003, Huang, et al.; 2004, Zuk, et al.; 2001). Under inductive conditions ASCs express aggrecan, chondroitin sulphate, collagen type II and IV and proteoglycans associated with chondrogenic phenotype.(Awad, et al.; 2004, Erickson, et al.; 2002, Wickham, et al.; 2003, Zuk, et al.; 2001)
For chondrogenic differentiation, ASCs cells are grown to confluency in 30-mm dishes and incubated for 3 weeks in DMEM containing 1% FBS, 50 mM L-ascorbic acid-2-phosphate, 40 mg/ml proline, 100 mg/ml pyruvate, 10 ng/ml transforming growth factor (Harriman, et al.)-b3, and 1x ITS. Induction medium is replaced every 3 days. At the indicated time points, differentiated cells are fixed for 1h with 4% paraformaldehyde and rinsed with PBS. Accumulation of chondrocyte matrix is detected with alcian blue staining (pH 2.5, Wako) (Matsumoto, et al.; 2008).
3.1.6. Neuronal differentiation
There is preliminary evidence suggesting that human ASCs can display neuronal and/or oligodendrocytic markers. ADSC at passages 2–5 are seeded in six-well plates at 40%–60%
confluence. After three washes with PBS, the cells are induced with NIM (DMEM supplemented with 500 mM IBMX, 200 mM INDO, and 5 mg/ml insulin) for 1 hr. The cells are then examined for the expression of neuronal markers S100, NF70, and nestin followed by hematoxylin-eosin (HE) staining (Ning, et al.; 2006).
The
4. Biomaterials in tissue engineering with ASCs
Currently, autologous and allogenic adipose tissues represent a ubiquitous source of material for fat reconstructive therapies. However, these approaches are limited, and often accompanied by a 40–60% reduction in graft volume following transplantation. A number of factors including a stable scaffold support structure and vascularisation is necessary to support
For functional tissue replacement such as bone, Silk-based biomaterials have previously been demonstrated to offer exceptional benefits over conventional synthetic (e.g. poly-glycolic and lactic acid copolymers) and natural (e.g. collagen type I) biomaterials (Meinel, et al.; 2004). The slow degradation and mechanical integrity of silk scaffolds in comparison with other conventional biomaterials such as collagen and PLA, above all for long-term
For
Tissue-specific scaffolds and signalling systems are essential to differentiate stem cells into the required cells and use them effectively to construct three-dimensional (3D) tissues (Sterodimas, et al.; 2010). It has also been proved that adipose tissues engineered with ASCs and type I collagen scaffolds can serve
Porous collagenous microbeads can be useful as injectable cell delivery vehicles for adipose-derived stem cells, allowing
Placental decellular matrix (PDM) holds potential as a scaffold for adipose tissue engineering applications. The placenta is a rich source of human extracellular matrix (ECM) components that can be harvested without harm to the donor. Constructs derived from the ECM may mimic the native environment of the body, promoting normal cellular organization and behavior. Natural materials also have advantages in terms of ease of processing, biodegradability and biocompatibility (Schmidt and Baier; 2000). Cell-adhesive placental decellular matrix scaffolds facilitate proliferation and viability, while differentiation is augmented when the cells are encapsulated in non-adhesive cross-linked hyaluronan (XLHA) scaffolds (Flynn, et al.; 2008). Incorporation of XLHA into the PDM scaffolds may improve the construct bulking properties and may influence cellular infiltration, differentiation and wound healing (Shu XZ; 2004).
Other candidate for suitable scaffolds is non-woven polyglycolic acid (PGA) and hyaluronic acid gel. In a recent study it has been shown that more adipose-tissue-like construct is regenerated when using type I collagen sponge than when the non-woven polyglycolic acid or hyaluronic acid gel are used (Itoi, et al.; 2010). In addition, significant evidence has been shown that ASCs and PLGA spheres can be used in a clinical setting to generate adipose tissue as a noninvasive soft tissue filler (Choi, et al.; 2006).
Altman et al. could show that human adipose-derived stem cells seeded on a silk fibroin-chitosan scaffold enhance wound healing and show differentiation into fibrovascular, endothelial, and epithelial components of restored tissue (Altman, et al.; 2009). In addition, it has been shown that transfection of human ASCs with liposome- enveloped xenogenic protein from a neonatal rat tissue preparation can induce differentiation of stem cells along the directed lineage (Gaustad, et al.; 2004). These observations support the hypothesis that the inductive biochemical and structural cues of the microenvironment are conserved across species and that a silk fibroin-chitosan delivery vehicle can provide a beneficial niche in supporting migration, proliferation, and differentiation of the applied cells (Altman, et al.; 2009).
Another complex biomolecule which has sparked great interest for tissue engineering is Hyaluronic acid which has been stated to support the growth and development of progenitor cells (Brun, et al.; 1999, Solchaga, et al.; 1999). The material has a progressive rate of biodegradation, lacks cytotoxicity and does not induce a systemic immune response or chronic inflammation in a human
Long-standing, 3D predefined-shape adipose tissue from hAD-MSCs of human adipose tissue remains a challenge. Lin et al. cultured scaffolds (Gelatin sponges, monofilament polypropylene and polyglycolic acid meshes) with hAD-MSCs in adipogenic medium for 2 weeks before implantation, and implanted scaffolds were harvested after 2, 4, and 6 months
There are numerous adipose tissue engineering culture strategies in which the core tissue engineering principles comprising appropriate cells, scaffold, and microenvironment are optimized (Patrick; 2001). Specifically, there are static versus dynamic culture, co-cultivation, and addition of growth factors, vascularization, and long-term sustainability of engineered constructs. The advantage of dynamic culture includes increased nutrient and oxygen delivery to cells within a 3D construct (Frye and Patrick; 2006). Improved bioreactor designs to address direct perfusion conditions are necessary to advance dynamic culture techniques (Choi, et al.; 2010).
It has been shown that hASCs express pericyte lineage markers
5. Conclusion
ASCs provide an abundant and readily accessible source of multipotent stem cells. The use of autologous stem cells expanded
However, further studies are needed before ASCs can be used clinically. In particular, investigators need to demonstrate the safety and efficacy of ASCs cells in animal models, either alone or in combination with novel biomaterial scaffolds.
Acknowledgments
Both the authors Dr. Mathias Tremp and Dr. Souzan Salemi contributed equivalently in writing this chapter.
References
- 1.
Ahn H. H. Lee J. H. Kim K. S. Lee J. Y. Kim M. S. Khang G. Lee I. W. Lee H. B. 2008 Polyethyleneimine-mediated gene delivery into human adipose derived stem cells.29 15 2415 2422 0142-9612 - 2.
Altman A. M. Yan Y. Matthias N. Bai X. Rios C. Mathur A. B. Song Y. H. Alt E. U. 2009 IFATS collection: Human adipose-derived stem cells seeded on a silk fibroin-chitosan scaffold enhance wound repair in a murine soft tissue injury model.27 1 250 258 1549-4918 - 3.
Amos P. J. Shang H. Bailey A. M. Taylor A. Katz A. J. Peirce S. M. 2008 IFATS collection: The role of human adipose-derived stromal cells in inflammatory microvascular remodeling and evidence of a perivascular phenotype.26 10 2682 2690 1549-4918 - 4.
Aoki T. Ohnishi H. Oda Y. Tadokoro M. Sasao M. Kato H. Hattori K. Ohgushi H. 2010 Generation of induced pluripotent stem cells from human adipose-derived stem cells without c-MYC.16 7 2197 2206 0193-7335 X - 5.
Aust L. Devlin B. Foster S. J. Halvorsen Y. D. Hicok K. du Laney. T. Sen A. Willingmyre G. D. Gimble J. M. 2004 Yield of human adipose-derived adult stem cells from liposuction aspirates.6 1 7 14 1465-3249 - 6.
Awad H. A. Halvorsen Y. D. Gimble J. M. Guilak F. 2003 Effects of transforming growth factor beta1 and dexamethasone on the growth and chondrogenic differentiation of adipose-derived stromal cells.9 6 1301 1312 1076-3279 - 7.
Awad H. A. Wickham M. Q. Leddy H. A. Gimble J. M. Guilak F. 2004 Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds.25 16 3211 3222 0142-9612 - 8.
Beamish J. A. He P. Kottke-Marchant K. Marchant R. E. 2010 Molecular regulation of contractile smooth muscle cell phenotype: implications for vascular tissue engineering. Part B Rev16 5 467 491 1937-3376 - 9.
Becker A. J. Mc C. E. Till J. E. 1963 Cytological demonstration of the clonal nature of spleen colonies derived from transplanted mouse marrow cells.197 452 454 0028-0836 - 10.
Borges J. Mueller M. C. Padron N. T. Tegtmeier F. Lang E. M. Stark G. B. 2003 Engineered adipose tissue supplied by functional microvessels.9 6 1263 1270 1076-3279 - 11.
Boubaker el Andalousi. R. Daussin P. A. Micallef J. P. Roux C. Nougues J. Chammas M. Reyne Y. Bacou F. 2002 Changes in mass and performance in rabbit muscles after muscle damage with or without transplantation of primary satellite cells.11 2 169 180 0963-6897 - 12.
Brun P. Cortivo R. Zavan B. Vecchiato N. Abatangelo G. 1999 In vitro reconstructed tissues on hyaluronan-based temporary scaffolding.10 10 683 688 0957-4530 - 13.
Choi J. H. Gimble J. M. Lee K. Marra K. G. Rubin J. P. Yoo J. J. Vunjak-Novakovic G. Kaplan D. L. 2010 Adipose tissue engineering for soft tissue regeneration. Part B Rev16 4 413 426 1937-3376 - 14.
Choi Y. S. Cha S. M. Lee Y. Y. Kwon S. W. Park C. J. Kim M. 2006 Adipogenic differentiation of adipose tissue derived adult stem cells in nude mouse.345 2 631 637 0000-6291 X - 15.
Cowherd R. M. Lyle R. E. Mc Gehee R. E. Jr 1999 Molecular regulation of adipocyte differentiation.10 1 3 10 1084-9521 - 16.
Crisan M. Yap S. Casteilla L. Chen C. W. Corselli M. Park T. S. Andriolo G. Sun B. Zheng B. Zhang L. Norotte C. Teng P. N. Traas J. Schugar R. Deasy B. M. Badylak S. Buhring H. J. Giacobino J. P. Lazzari L. Huard J. Peault B. 2008 A perivascular origin for mesenchymal stem cells in multiple human organs.3 3 301 313 1875-9777 - 17.
De Rosa A. De Francesco F. Tirino V. Ferraro G. A. Desiderio V. Paino F. Pirozzi G. D’Andrea F. Papaccio G. 2009 A new method for cryopreserving adipose-derived stem cells: an attractive and suitable large-scale and long-term cell banking technology.15 4 659 667 1937-3392 - 18.
De Ugarte D. A. Morizono K. Elbarbary A. Alfonso Z. Zuk P. A. Zhu M. Dragoo J. L. Ashjian P. Thomas B. Benhaim P. Chen I. Fraser J. Hedrick M. H. 2003 Comparison of multi-lineage cells from human adipose tissue and bone marrow.174 3 101 109 1422-6405 - 19.
Dellavalle A. Sampaolesi M. Tonlorenzi R. Tagliafico E. Sacchetti B. Perani L. Innocenzi A. Galvez B. G. Messina G. Morosetti R. Li S. Belicchi M. Peretti G. Chamberlain J. S. Wright W. E. Torrente Y. Ferrari S. Bianco P. Cossu G. 2007 Pericytes of human skeletal muscle are myogenic precursors distinct from satellite cells.9 3 255 267 1465-7392 - 20.
Deslex S. Negrel R. Vannier C. Etienne J. Ailhaud G. 1987 Differentiation of human adipocyte precursors in a chemically defined serum-free medium.11 1 19 27 0307-0565 - 21.
Dragoo J. L. Samimi B. Zhu M. Hame S. L. Thomas B. J. Lieberman J. R. Hedrick M. H. Benhaim P. 2003 Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads. Br85 5 740 747 0030-1620 X - 22.
Engfeldt P. Arner P. Ostman J. 1980 Influence of adipocyte isolation by collagenase on phosphodiesterase activity and lipolysis in man.21 4 443 448 0022-2275 - 23.
Erickson G. R. Gimble J. M. Franklin D. M. Rice H. E. Awad H. Guilak F. 2002 Chondrogenic potential of adipose tissue-derived stromal cells in vitro and in vivo.290 2 763 769 0000-6291 X - 24.
Estes B. T. Diekman B. O. Guilak F. 2008 Monolayer cell expansion conditions affect the chondrogenic potential of adipose-derived stem cells.99 4 986 995 1097-0290 - 25.
Farmer S. R. 2006 Transcriptional control of adipocyte formation.4 4 263 273 1550-4131 - 26.
Flynn L. E. Prestwich G. D. Semple J. L. Woodhouse K. A. 2008 Proliferation and differentiation of adipose-derived stem cells on naturally derived scaffolds.29 12 1862 1871 0142-9612 - 27.
Frye C. A. Patrick C. W. 2006 Three-dimensional adipose tissue model using low shear bioreactors.42 5-6 109 114 1071-2690 - 28.
Gaustad K. G. Boquest A. C. Anderson B. E. Gerdes A. M. Collas P. 2004 Differentiation of human adipose tissue stem cells using extracts of rat cardiomyocytes.314 2 420 427 0000-6291 X - 29.
Gimble J. Guilak F. 2003 Adipose-derived adult stem cells: isolation, characterization, and differentiation potential.5 5 362 369 1465-3249 - 30.
Gimble J. M. Katz A. J. Bunnell B. A. 2007 Adipose-derived stem cells for regenerative medicine.100 9 1249 1260 1524-4571 - 31.
Gonda K. Shigeura T. Sato T. Matsumoto D. Suga H. Inoue K. Aoi N. Kato H. Sato K. Murase S. Koshima I. Yoshimura K. 2008 Preserved proliferative capacity and multipotency of human adipose-derived stem cells after long-term cryopreservation.121 2 401 410 1529-4242 - 32.
Gregoire F. M. Smas C. M. Sul H. S. 1998 Understanding adipocyte differentiation.78 3 783 809 0031-9333 - 33.
Gronthos S. Franklin D. M. Leddy H. A. Robey P. G. Storms R. W. Gimble J. M. 2001 Surface protein characterization of human adipose tissue-derived stromal cells.189 1 54 63 0021-9541 - 34.
Guilak F. Estes B. T. Diekman B. O. Moutos F. T. Gimble J. M. 2010 Nicolas Andry Award: Multipotent adult stem cells from adipose tissue for musculoskeletal tissue engineering. Res468 9 2530 2540 1528-1132 - 35.
Halvorsen Y. D. Bond A. Sen A. Franklin D. M. Lea-Currie Y. R. Sujkowski D. Ellis P. N. Wilkison W. O. Gimble J. M. 2001 Thiazolidinediones and glucocorticoids synergistically induce differentiation of human adipose tissue stromal cells: biochemical, cellular, and molecular analysis.50 4 407 413 0026-0495 - 36.
Halvorsen Y. D. Franklin D. Bond A. L. Hitt D. C. Auchter C. Boskey A. L. Paschalis E. P. Wilkison W. O. Gimble J. M. 2001 Extracellular matrix mineralization and osteoblast gene expression by human adipose tissue-derived stromal cells.7 6 729 741 1076-3279 - 37.
Harriman G. R. Bogue M. Rogers P. Finegold M. Pacheco S. Bradley A. Zhang Y. Mbawuike I. N. 1999 Targeted deletion of the IgA constant region in mice leads to IgA deficiency with alterations in expression of other Ig isotypes.162 5 2521 2529 0022-1767 - 38.
Hauner H. Entenmann G. Wabitsch M. Gaillard D. Ailhaud G. Negrel R. Pfeiffer E. F. 1989 Promoting effect of glucocorticoids on the differentiation of human adipocyte precursor cells cultured in a chemically defined medium.84 5 1663 1670 0021-9738 - 39.
Hemmrich K. von Heimburg. D. Rendchen R. Di Bartolo C. Milella E. Pallua N. 2005 Implantation of preadipocyte-loaded hyaluronic acid-based scaffolds into nude mice to evaluate potential for soft tissue engineering.26 34 7025 7037 0142-9612 - 40.
Hicok K. C. Du Laney. T. V. Zhou Y. S. Halvorsen Y. D. Hitt D. C. Cooper L. F. Gimble J. M. 2004 Human adipose-derived adult stem cells produce osteoid in vivo.10 3-4 371 380 1076-3279 - 41.
Ho J. H. Ma W. H. Tseng T. C. Chen Y. F. Chen M. H. Lee O. K. 2010 Isolation and Characterization of Multi-Potent Stem Cells from Human Orbital Fat Tissues. Part A, pp.,0193-7335 1937 335 X - 42.
Huang J. I. Zuk P. A. Jones N. F. Zhu M. Lorenz H. P. Hedrick M. H. Benhaim P. 2004 Chondrogenic potential of multipotential cells from human adipose tissue.113 2 585 594 0032-1052 - 43.
Itoi Y. Takatori M. Hyakusoku H. Mizuno H. 2010 Comparison of readily available scaffolds for adipose tissue engineering using adipose-derived stem cells.63 5 858 864 1878-0539 - 44.
Jack G. S. Zhang R. Lee M. Xu Y. Wu B. M. Rodriguez L. V. 2009 Urinary bladder smooth muscle engineered from adipose stem cells and a three dimensional synthetic composite.30 19 3259 3270 1878-5905 - 45.
Kakudo N. Minakata T. Mitsui T. Kushida S. Notodihardjo F. Z. Kusumoto K. 2008 Proliferation-promoting effect of platelet-rich plasma on human adipose-derived stem cells and human dermal fibroblasts.122 5 1352 1360 1529-4242 - 46.
Kang S. K. Lee D. H. Bae Y. C. Kim H. K. Baik S. Y. Jung J. S. 2003 Improvement of neurological deficits by intracerebral transplantation of human adipose tissue-derived stromal cells after cerebral ischemia in rats.183 2 355 366 0014-4886 - 47.
Katz A. J. Tholpady A. Tholpady S. S. Shang H. Ogle R. C. 2005 Cell surface and transcriptional characterization of human adipose-derived adherent stromal (hADAS) cells.23 3 412 423 1066-5099 - 48.
Kirkland J. L. Hollenberg C. H. 1998 Inhibitors of preadipocyte replication: opportunities for the treatment of obesity.20 177 195 0079-6484 - 49.
Lazar M. A. 2005 PPAR gamma, 10 years later.87 1 9 13 0300-9084 - 50.
Lee J. A. Parrett B. M. Conejero J. A. Laser J. Chen J. Kogon A. J. Nanda D. Grant R. T. Breitbart A. S. 2003 Biological alchemy: engineering bone and fat from fat-derived stem cells.50 6 610 617 0148-7043 - 51.
Lendeckel S. Jodicke A. Christophis P. Heidinger K. Wolff J. Fraser J. K. Hedrick M. H. Berthold L. Howaldt H. P. 2004 Autologous stem cells (adipose) and fibrin glue used to treat widespread traumatic calvarial defects: case report.32 6 370 373 1010-5182 - 52.
Lin S. D. Wang K. H. Kao A. P. 2008 Engineered adipose tissue of predefined shape and dimensions from human adipose-derived mesenchymal stem cells. Part A14 5 571 581 1937-3341 - 53.
Liu Z. J. Zhuge Y. Velazquez O. C. 2009 Trafficking and differentiation of mesenchymal stem cells.106 6 984 991 1097-4644 - 54.
Lu F. Gao J. H. Ogawa R. Mizuro H. Hykusoku H. 2006 Adipose tissues differentiated by adipose-derived stem cells harvested from transgenic mice.9 6 359 364 1008-1275 - 55.
Lu F. Mizuno H. Uysal C. A. Cai X. Ogawa R. Hyakusoku H. 2008 Improved viability of random pattern skin flaps through the use of adipose-derived stem cells.121 1 50 58 1529-4242 - 56.
Matsumoto T. Kano K. Kondo D. Fukuda N. Iribe Y. Tanaka N. Matsubara Y. Sakuma T. Satomi A. Otaki M. Ryu J. Mugishima H. 2008 Mature adipocyte-derived dedifferentiated fat cells exhibit multilineage potential.215 1 210 222 1097-4652 - 57.
Mauney J. R. Nguyen T. Gillen K. Kirker-Head C. Gimble J. M. Kaplan D. L. 2007 Engineering adipose-like tissue in vitro and in vivo utilizing human bone marrow and adipose-derived mesenchymal stem cells with silk fibroin 3D scaffolds.28 35 5280 5290 0142-9612 - 58.
Meinel L. Karageorgiou V. Hofmann S. Fajardo R. Snyder B. Li C. Zichner L. Langer R. Vunjak-Novakovic G. Kaplan D. L. 2004 Engineering bone-like tissue in vitro using human bone marrow stem cells and silk scaffolds. A71 1 25 34 1549-3296 - 59.
Mizuno H. Zuk P. A. Zhu M. Lorenz H. P. Benhaim P. Hedrick M. H. 2002 Myogenic differentiation by human processed lipoaspirate cells.109 1 199 209 discussion 210-1,0032-1052 - 60.
Moseley T. A. Zhu M. Hedrick M. H. 2006 Adipose-derived stem and progenitor cells as fillers in plastic and reconstructive surgery.118 3 Suppl,121S 128S 1529-4242 - 61.
Nam S. Y. Lobie P. E. 2000 The mechanism of effect of growth hormone on preadipocyte and adipocyte function.1 2 73 86 1467-7881 - 62.
Ning H. Lin G. Lue T. F. Lin C. S. 2006 Neuron-like differentiation of adipose tissue-derived stromal cells and vascular smooth muscle cells.74 9-10 510 518 0301-4681 - 63.
Ogawa R. Mizuno H. Watanabe A. Migita M. Hyakusoku H. Shimada T. 2004 Adipogenic differentiation by adipose-derived stem cells harvested from GFP transgenic mice-including relationship of sex differences.319 2 511 517 0000-6291 X - 64.
Owens G. K. Kumar M. S. Wamhoff B. R. 2004 Molecular regulation of vascular smooth muscle cell differentiation in development and disease.84 3 767 801 0031-9333 - 65.
Parker A. M. Shang H. Khurgel M. Katz A. J. 2007 Low serum and serum-free culture of multipotential human adipose stem cells.9 7 637 646 1465-3249 - 66.
Patrick C. W. Jr 2001 Tissue engineering strategies for adipose tissue repair.263 4 361 366 0000-3276 X - 67.
Patrick C. W. Jr Chauvin P. B. Hobley J. Reece G. P. 1999 Preadipocyte seeded PLGA scaffolds for adipose tissue engineering.5 2 139 151 1076-3279 - 68.
Patrick C. W. Jr Zheng B. Johnston C. Reece G. P. 2002 Long-term implantation of preadipocyte-seeded PLGA scaffolds.8 2 283 293 1076-3279 - 69.
Pittenger M. F. Mackay A. M. Beck S. C. Jaiswal R. K. Douglas R. Mosca J. D. Moorman M. A. Simonetti D. W. Craig S. Marshak D. R. 1999 Multilineage potential of adult human mesenchymal stem cells.284 5411 143 147 0036-8075 - 70.
Preece A. 1972 A Manual for Histologic Technicians. Boston, MA: Little, Brown and Co. - 71.
Rodbell M. 1966 Metabolism of isolated fat cells. II. The similar effects of phospholipase C (Clostridium perfringens alpha toxin) and of insulin on glucose and amino acid metabolism.241 1 130 139 0021-9258 - 72.
Rodbell M. 1966 The metabolism of isolated fat cells. IV. Regulation of release of protein by lipolytic hormones and insulin.241 17 3909 3917 0021-9258 - 73.
Rodbell M. Jones A. B. 1966 Metabolism of isolated fat cells. 3. The similar inhibitory action of phospholipase C (Clostridium perfringens alpha toxin) and of insulin on lipolysis stimulated by lipolytic hormones and theophylline.241 1 140 142 0021-9258 - 74.
Rubin J. P. Bennett J. M. Doctor J. S. Tebbets B. M. Marra K. G. 2007 Collagenous microbeads as a scaffold for tissue engineering with adipose-derived stem cells.120 2 414 424 1529-4242 - 75.
Sakuma T. Matsumoto T. Kano K. Fukuda N. Obinata D. Yamaguchi K. Yoshida T. Takahashi S. Mugishima H. 2009 Mature, adipocyte derived, dedifferentiated fat cells can differentiate into smooth muscle-like cells and contribute to bladder tissue regeneration.182 1 355 365 1527-3792 - 76.
Schmidt C. E. Baier J. M. 2000 Acellular vascular tissues: natural biomaterials for tissue repair and tissue engineering.21 22 2215 2231 0142-9612 - 77.
Sen A. Lea-Currie Y. R. Sujkowska D. Franklin D. M. Wilkison W. O. Halvorsen Y. D. Gimble J. M. 2001 Adipogenic potential of human adipose derived stromal cells from multiple donors is heterogeneous.81 2 312 319 0730-2312 - 78.
Shanahan C. M. Weissberg P. L. Metcalfe J. C. 1993 Isolation of gene markers of differentiated and proliferating vascular smooth muscle cells.73 1 193 204 0009-7330 - 79.
Shillabeer G. Forden J. M. Lau D. C. 1989 Induction of preadipocyte differentiation by mature fat cells in the rat.84 2 381 387 0021-9738 - 80.
Shu X. Z. P. G. 2004 Therapeutic biomaterials from chemically modified hyaluronan. In: Garg HG, Hales CA, editors . Amsterdam: Elsevier Press,475 504 - 81.
Silberstein L. Webster S. G. Travis M. Blau H. M. 1986 Developmental progression of myosin gene expression in cultured muscle cells.46 7 1075 1081 0092-8674 - 82.
Solchaga L. A. Dennis J. E. Goldberg V. M. Caplan A. I. 1999 Hyaluronic acid-based polymers as cell carriers for tissue-engineered repair of bone and cartilage.17 2 205 213 0736-0266 - 83.
Sorisky A. 1999 From preadipocyte to adipocyte: differentiation-directed signals of insulin from the cell surface to the nucleus.36 1 1 34 1040-8363 - 84.
Sterodimas A. de Faria J. Nicaretta B. Pitanguy I. 2010 Tissue engineering with adipose-derived stem cells (ADSCs): current and future applications.63 11 1886 1892 1878-0539 - 85.
Stillaert F. B. Di Bartolo C. Hunt J. A. Rhodes N. P. Tognana E. Monstrey S. Blondeel P. N. 2008 Human clinical experience with adipose precursor cells seeded on hyaluronic acid-based spongy scaffolds.29 29 3953 3959 0142-9612 - 86.
Tchkonia T. Giorgadze N. Pirtskhalava T. Tchoukalova Y. Karagiannides I. Forse R. A. De Ponte M. Stevenson M. Guo W. Han J. Waloga G. Lash T. L. Jensen M. D. Kirkland J. L. 2002 Fat depot origin affects adipogenesis in primary cultured and cloned human preadipocytes.282 5 R1286 R1296 0363-6119 - 87.
Toyoda M. Matsubara Y. Lin K. Sugimachi K. Furue M. 2009 Characterization and comparison of adipose tissue-derived cells from human subcutaneous and omental adipose tissues.27 7 440 447 1099-0844 - 88.
von Heimburg. D. Zachariah S. Heschel I. Kuhling H. Schoof H. Hafemann B. Pallua N. 2001 Human preadipocytes seeded on freeze-dried collagen scaffolds investigated in vitro and in vivo.22 5 429 438 0142-9612 - 89.
von Heimburg. D. Zachariah S. Low A. Pallua N. 2001 Influence of different biodegradable carriers on the in vivo behavior of human adipose precursor cells.108 2 411 420 discussion 421-2,0032-1052 - 90.
Weintraub H. Davis R. Tapscott S. Thayer M. Krause M. Benezra R. Blackwell T. K. Turner D. Rupp R. Hollenberg S. et al. 1991 The myoD gene family: nodal point during specification of the muscle cell lineage.251 4995 761 766 0036-8075 - 91.
Wickham M. Q. Erickson G. R. Gimble J. M. Vail T. P. Guilak F. 2003 Multipotent stromal cells derived from the infrapatellar fat pad of the knee. ,412 196 212 0000-9921 X - 92.
Yoshimura K. Sato K. Aoi N. Kurita M. Hirohi T. Harii K. 2008 Cell-assisted lipotransfer for cosmetic breast augmentation: supportive use of adipose-derived stem/stromal cells.32 1 48 55 discussion 56-7,0036-4216 X - 93.
Yuan Q. Zeng X. Chen L. Peng E. Ye Z. 2010 Comparison of myogenic differentiation ability of adipose-derived stem cells from different sites in rabbit]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi24 10 1228 1232 1002-1892 - 94.
Zhang J. Shehabeldin A. da Cruz. L. A. Butler J. Somani A. K. Mc Gavin M. Kozieradzki I. dos Santos. A. O. Nagy A. Grinstein S. Penninger J. M. Siminovitch K. A. 1999 Antigen receptor-induced activation and cytoskeletal rearrangement are impaired in Wiskott-Aldrich syndrome protein-deficient lymphocytes.190 9 1329 1342 0022-1007 - 95.
Zhang Y. S. Gao J. H. Lu F. Zhu M. Liao Y. J. 2007 Cellular compatibility of type collagen I scaffold and human adipose-derived stem cells].27 2 223 225 1673-4254 - 96.
Zuk P. A. Zhu M. Ashjian P. De Ugarte D. A. Huang J. I. Mizuno H. Alfonso Z. C. Fraser J. K. Benhaim P. Hedrick M. H. 2002 Human adipose tissue is a source of multipotent stem cells.13 12 4279 4295 1059-1524 - 97.
Zuk P. A. Zhu M. Mizuno H. Huang J. Futrell J. W. Katz A. J. Benhaim P. Lorenz H. P. Hedrick M. H. 2001 Multilineage cells from human adipose tissue: implications for cell-based therapies.7 2 211 228 1076-3279