The cartilage of joints is long‐lasting (i.e., permanent) cartilage and is not spontaneously repaired after injury in humans. There has been considerable interest in the clinical application of stem cells to the repair of damaged cartilage; however, current cell therapies using adult chondrocytes and mesenchymal stromal cells face problems associated with the low yield of such cells. The expansion culture, needed before transplantation, leads to the formation of fibrocartilage or growth plate-like (i.e., bone‐forming) cartilage in vivo. Both types of cartilage are unsuitable for the repair of joint cartilage such as meniscus and articular cartilage. Joints are formed during embryogenesis. Therefore, we hypothesize that embryonic progenitor cells responsible for the development of joint cartilage would be the best for regenerating joint cartilage in the adult. Pluripotent stem cells (PSCs) are expected to differentiate in culture into any somatic cell types through processes that mimic embryogenesis, making human (h)PSCs a promising source of embryonic cells for regenerative medicine. However, regardless of the cell system used, the major research goals leading to clinical application to cartilage regeneration are to (1) expand chondrogenic cells (chondroprogenitors) to sufficient numbers without loss of their chondrogenic activity, and (2) direct the differentiation of such cells in vivo or in vitro toward articular or other types of chondrocytes of interest. The overall aim of the current review was to provide the basis of a strategy for meeting the goals for cartilage regeneration by the use of hPSC‐derived chondroprogenitor cells. We provide an overview on signaling mechanisms that are known to affect the expandability and chondrogenic activity of adult and embryonic chondroprogenitors, as well as their differentiation in vivo or in vitro toward a particular type of chondrocyte. We then discuss alternative types of progenitor cells that might replace or combine with the hPSC‐derived chondroprogenitors to regenerate permanent cartilage. We also include our recent achievement of successfully expanding hPSC‐derived neural crest to generate ectomesenchymal chondroprogenitors that can be maintained for a long term in culture without loss of chondrogenic activity. Finally, we provide information on the challenges that hPSC progeny‐based regenerative medicine will face, and discuss the implications for such challenges for the future use of PSC progeny to regenerate cartilage.
Damaged human joint articular cartilage does not heal spontaneously and eventually develops into osteoarthritis, probably due to a lack of proper “regenerative (i.e., stem/progenitor)” cells. Consequently, there has been considerable interest in the clinical application of chondrogenic stem/progenitor cells in the repair of damaged cartilage. For example, cell‐based therapies have been developed that use the endogenous mobilization of marrow cells (i.e., microfracture method), or cells such as expanded (dedifferentiated) articular chondrocytes or mesenchymal stromal cells (MSCs) added exogenously via a periosteal patch or collagen membrane (i.e., autologous chondrocyte implantation method). Although such therapies relieve the major clinical symptom of pain, they do not promote the regeneration of long‐lasting hyaline cartilage. Tissue engineering approaches that deliver a biodegradable matrix seeded with dedifferentiated chondrocytes or MSCs have been tested for the treatment of large cartilage lesions in animal models with similarly disappointing long‐term results
As to the cell type to be used for cartilage regeneration, chondrocytes are naturally suitable. However, their use requires
Joint formation is initiated during embryogenesis by a specialized cell type, the “joint progenitor,” which is responsible for generating hyaline articular cartilage. Furthermore, surface injury that is introduced
Concerning the challenges with the use of MSCs as stated by Somoza et al. , it is conceivable that the successful repair of articular cartilage will depend on both the ability of chondrogenic cells with high expansion capacity to form hyaline cartilage and a microenvironment at the site of repair that promotes chondrogenesis, while preventing terminal maturation and mineralization of chondrocytes. Therefore, we aim to give an overview on what we know from genetic as well as cellular (e.g., MSCs and MDSCs) studies, (1) about ways to expand chondroprogenitors and maintain their chondrogenic activity, and (2) about the potential signaling mechanisms and cell populations that affect the type (e.g., articular chondrocytes or growth plate chondrocytes) and/or differentiation state (e.g., immature or mature) of chondrocytes generated from chondroprogenitors. We will also summarize relevant information obtained from our studies of hPSC‐derived chondroprogenitors.
2. Long‐term expansion of chondroprogenitor cells without loss of their chondrogenic activity: advantages of hPSC‐derived progenitors
With any cell system, one of the major research goals for cellular therapy or tissue‐engineering approach for cartilage regeneration is to expand chondroprogenitors to yield large numbers without the loss of chondrogenic activity. These goals are important because the quality and durability of the
2.1. FGF, TGFβ, hedgehog, and WNTs for the expansion of adult MSC
FGF signaling is known to upregulate expression of
Indian hedgehog (Ihh) is a key mitogen for chondrocytes in the growth plate . Similarly, hedgehog signaling stimulates the growth of human bone marrow MSCs and enhances chondrogenesis in serum‐containing culture conditions . On the other hand, WNT signaling is known to play important roles on the growth, specification, movement, and organization of early precursor cells for osteochondrogenesis . The roles of WNT signaling on the proliferation of embryonic osteochondrogenic mesenchymal cells  and their association with osteogenesis have been well established [48, 49]. Very recently, WNT3a has been shown to further stimulate the FGF2‐stimulated growth of hMSCs and enhance the FGF2‐enhanced maintenance of chondrogenic potency for at least 20 days (4 passages) of expansion culture in the presence of 10% FBS . In spite of these positive results with hedgehogs and WNTs in expanding mesenchymal cells, both factors are “developmental factors” that induce osteogenesis rather than chondrogenesis during skeletogenesis. Through the canonical signaling pathway (involving β‐catenin), WNTs reduce Sox9 levels and antagonize the functions of Sox9 in chondrocytes , and Ihh is placed upstream of the WNT/β‐catenin signaling events, essential for osteoblastogenesis in the perichondrium of developing bone [52–55]. We have also found that WNTs stimulate the growth of hPSC‐derived chondroprogenitor cells (data not shown). Although the effects may depend on the developmental/differentiation stage of the target cell, use of WNTs and Ihh on chondrogenic mesenchymal cells probably needs careful prior examination to verify that they do not cause loss of chondrogenic activity during expansion. Research into ways of improving expansion culture for hMSCs to preserve their chondrogenicity reproducibly is still ongoing.
2.2. Suppression of TGFβ signaling successfully expands endothelial (progenitor) cells derived from PSCs
Mesenchymal progeny of PSCs had the similar problems as adult MSC, namely the loss of original phenotypes and developmental potentials during expansion. In this regard, Miyazono and colleagues  published a pioneering work in 2003 in which they demonstrated that Nodal/Activin/TGFβ receptor kinase inhibitor, SB431542, enhanced the growth and integrity of mouse (m)ESC‐derived endothelial cells, leading to a successful protocol for the expansion of hPSC‐derived endothelial cells . TGFβ is known to be a potent inhibitor for lymphohematopoietic progenitor cell proliferation . However, as the name “transforming growth factor” indicates, it was originally found as a potent growth‐promoting factor for many transformed (tumor/cancer) cells and untransformed (normal) cells, including human adult bone marrow MSCs . PSC‐derived endothelial cells may produce TGFβ‐like activity that serves as a “brake” on their own proliferation in culture.
2.3. Long‐term expansion of endoderm stem/progenitor cells derived from hPSCs without loss of their developmental potency
Recently, multipotential endodermal stem/progenitor cells, which can undergo long‐term expansion without loss of their phenotypes and developmental potentials (i.e., can self‐renew), have been developed from hPSCs [59, 60]. The foregut stem cells can be maintained in culture over 18 passages in RPMI + B27‐based serum‐free medium supplemented with FGF2, Activin A, BMP4, hepatocyte growth factor (HGF), and epidermal growth factor (EGF) . The endodermal progenitor cells can be maintained over 24 passages in IMDM:F12 (3:1) + N2/B27‐based serum‐free medium containing FGF2, BMP4, VEGF, and EGF . Nodal/Activin/TGFβ receptor kinase inhibitor is not necessary for the long‐term expansion of endodermal stem/progenitor cells. Therefore, the inhibition of Nodal/Activin/TGFβ receptor kinase is not a common requirement for the expansion of hPSC‐derived progeny.
2.4. Long‐term maintenance of hPSC‐derived chondrogenic ectomesenchymal cells without loss of their chondrogenic activity
In contrast, methods for the long‐term expansion of hPSC‐derived chondrogenic mesenchymal cells had not been explored extensively. Research has focused more on the “genesis” but not “expansion” of chondrogenic activity from hPSCs. Although a short‐term expansion of chondrogenic activity as judged by the increase in the SOX9+ cell number was reported , expansion of PSC‐derived mesenchymal cells has not been accompanied by long‐term maintenance of their chondrogenic activity in the conventional MSC medium, as in the case of adult MSCs . Proliferation without loss of chondrogenic activity is thus not an intrinsic property of the hPSC‐derived “embryonic” chondrogenic cells. Rather, it is a property that can emerge under certain defined culture conditions. In this respect, we have previously established and refined signaling requirements for the differentiation of mouse and human ESCs and iPSCs to chondrogenic lateral plate mesoderm, paraxial mesoderm, and cranial neural crest‐like progeny in a serum‐free defined medium [22, 31, 33–35, 62–64]. Our group was the first to define the MIXL1‐green fluorescent protein (GFP)+ VEGFR2 (KDR)- PDGF receptor (PDGFR)α+ human paraxial mesoderm progeny and T‐GFP+KDR-PDGFRα+ mouse paraxial mesoderm progeny, from which chondrogenic mesenchymal cells were derived [33–35, 62]. We have also recently reported that MIXL1‐GFP-CD271hiCD73- human neural crest‐like progeny develops in a chemically defined medium [CDM: IMDM:F12 (1:1), 0.5% (w/v) fatty acid‐free BSA, 1% (v/v) synthetic lipids, 10 μg/ml insulin, 300 μg/ml holo‐transferrin, 0.17 mM ascorbic acid‐2‐phosphate, 0.3 mM monothyoglycerol] in the presence of Nodal/Activin/TGFβ receptor inhibitor (SB431542) [65–68] and that they quickly become CD271+CD73+ ectomesenchymal cells expressing
In contrast to PSC‐derived endothelial cells and chondrogenic ectomesenchymal cells, the maintenance of human adult bone marrow MSCs in culture required TGFβ in a defined culture conditions . Therefore, PSC‐derived ectomesenchymal cells and bone marrow MSCs are distinct cell types, despite the proposal that both may share the common developmental origin of neural crest [69–71]. In support, application of SB431542 to mouse bone marrow MSC culture failed to support their growth and preferentially left adipocytes (data not shown), consistent with the notion that TGFβ suppresses adipocytic differentiation of mesenchymal progenitors .
3. Signaling mechanisms potentially leading to the genesis of long‐lasting immature cartilage from hPSC‐derived chondroprogenitor cells
The reproducible generation, either
3.1. Indian hedgehiog (Ihh)‐parathyroid hormone‐related peptide (PTHrP) feedback loop
The Ihh‐PTHrP feedback loop is one of the best‐known mechanisms that controls chondrocyte hypertrophic differentiation in the growth plate [86–89]. After the cartilage primordium has formed during embryogenesis, perichondrial cells and chondrocytes at the ends of the cartilage produce PTHrP that signals through PTH receptors on chondrocytes. PTHrP action keeps chondrocytes in a proliferative state and delays their differentiation toward the post‐mitotic hypertrophic chondrocytes . The chondrocytes distant from the cartilage ends escape from such PTHrP effects and stop proliferating (pre‐hypertrophic chondrocytes). Such chondrocytes synthesize Ihh, which in turn stimulates the synthesis of PTHrP from chondrocytes at the ends of the cartilage, potentially through the action of TGFβ [91, 92]. The pre‐hypertrophic chondrocytes differentiate to hypertrophic chondrocytes and then to terminally matured, mineralized chondrocytes. The feedback loop between PTHrP and Ihh thus regulates the pace of differentiation of immature (proliferating) chondrocytes. The Ihh‐PTHrP system may also function similarly during articular chondrogenesis .
Elucidation of the individual roles of hedgehog and PTHrP signaling is important if there is to be future application of the Ihh and PTHrP signaling mechanisms to control chondrocyte hypertrophic differentiation during chondrogenesis from MSCs‐ or hPSC‐derived chondroprogenitors. To date, the role of hedgehog signaling on chondrocyte maturation appears contradictory. Ihh directly stimulates proliferation of (immature) chondrocyte but also stimulates osteogenesis from osteochondrogenic mesenchymal cells. In addition, in the absence of PTHrP, Ihh appears to stimulate hypertrophic differentiation of chondrocytes through activating canonical WNT signaling (analogous to the Ihh‐induced osteogenesis process) [94, 95] and vascularization of the hypertrophic cartilage, leading to trabecular bone formation [44, 96]. However, the overproduction of Sonic hedgehog (Shh) in cartilage seems to upregulate Sox9 directly in chondrocytes and chondrogenic mesenchymal cells and prevents or slows the maturation of chondrocytes [97, 98].
In contrast, the effect of PTHrP on the suppression of chondrocyte hypertrophic differentiation seems clear. The critical mechanism of PTHrP in keeping chondrocytes proliferative and delaying their hypertrophic differentiation is the activation of Gs and adenylyl cyclase to increase the level of intracellular cyclic adenosine monophosphate (cAMP) . The rise in cAMP leads to the suppression of Kip2 cyclin inhibitor expression  and activation of Sox9 by cAMP‐dependent protein kinase (PKA)‐mediated phosphorylation [101, 102]. PTHrP decreases the level of
Investigation of the effects of PTHrP on
3.2. TGFβ‐BMP signaling
TGFβ was first identified as cartilage‐inducing factor (CIF‐A) purified from bovine demineralized bone  but was soon found to have a negative effect on chondrocyte terminal maturation during scaffold‐free 3‐dimensional (3D) chondrogenesis culture (called “pellet culture”) . In fact, canonical TGFβ signaling that goes through the TGFβ receptor (TGFBR)–Smads such as Smad3 inhibits chondrocyte hypertrophic differentiation in the growth plate and, interestingly, also during
Thus, one way to stimulate chondrogenesis from chondroprogenitor cells while preventing hypertrophic differentiation of developed chondrocytes is to avoid overt activation of BMP signaling. Another is to select the appropriate BMP to use. For example, GDF5, one of the BMP‐family proteins isolated from articular cartilage [139, 140], is involved in joint morphogenesis in the mouse [139, 141–145]. GDF5 is also needed for proper maintenance of articular cartilage in adult humans [146–149].
Therefore, control of the strength and temporal action of BMP signaling via exogenously added BMPs such as GDF5 or BMP7 or by inhibitors for ALK1 or other BMPR–Smad activating receptors during chondrogenesis may lead to an optimal condition for hPSC‐derived chondroprogenitors to form permanent cartilage preferentially. Alternatively, combinatory control with other signaling mechanisms such as WNT signaling may be needed.
3.3. WNT signaling
As noted, WNT signaling through the canonical pathway of stabilization and nuclear translocation of β‐catenin stimulates osteogenesis and inhibits chondrogenesis by blocking
On the other hand, WNTs are implicated in the commitment of MSCs to differentiation. For example, WNT signals promote osteogenesis from MSCs by inducing
3.4. Natriuretic peptide signaling
The C‐type natriuretic peptide (CNP) signaling pathway is a major contributor to postnatal skeletal growth in humans [168, 169]. The CNP gene
3.5. VEGF signaling
For tissue engineering approaches to cartilage repair, extensive improvements have been made to scaffold/hydrogel technology to provide a suitable chemical and physical environment for the embedded chondrocytes or chondroprogenitors. However, the growth factors tested to date have been selected for their potential to facilitate chondrogenesis (e.g., TGFβs or BMPs) rather than to prevent terminal maturation and mineralization. VEGF, the best known angiogenic factor, is produced in the growth plate specifically from hypertrophic chondrocytes, not from the resting or proliferating immature chondrocytes. Furthermore, VEGF‐mediated blood vessel invasion is essential for coupling resorption of cartilage to bone formation . Since the VEGFR2 (KDR/Flk1) is also expressed in hypertrophic chondrocytes, it is expected that the suppression of VEGF function may prevent terminal maturation and mineralization. The VEGF inhibition strategy has already been applied to stem cell‐based cartilage regeneration therapy in animal models. Orthotopically transplanted MDSCs require BMP4 to differentiate effectively into chondrocytes to repair osteochondral defects in articular cartilage . However, the suppression of VEGF function by forced expression of a soluble form of VEGFR1 (sFlt1) as well as BMP4 significantly enhanced the repair function of MDSCs [3, 5]. Incorporation of an inhibitor of VEGF signaling in a scafford embedded with human nasal chondrocytes allowed
Bony cartilage recovered from transplanted mice, which originates from cartilage pellets generated by the hPSC‐derived ectomesenchymal cells (Figure 3 EctM), is usually highly vascular and becomes larger than the transplanted pellet (data not shown). In contrast, unmineralized or partially mineralized cartilage, which originates from cartilage pellets generated by uncultured paraxial mesoderm (Figure 3 PM), is generally not vascular and slightly smaller than the transplanted pellet (data not shown). Suppression of production or function of VEGF from chondrocytes developed from adult‐ or hPSC‐derived chondroprogenitors may thus lead to long‐lasting (i.e., permanent) cartilage
4. Cell types potentially leading to the genesis of permanent cartilage
The manipulation of signaling mechanisms may not be sufficient to generate permanent articular cartilage reproducibly
4.1. Nasal chondrocytes and their precursors (ectomesenchymal cells)‐novel chondroprogenitor cells
Numerous attempts to regenerate permanent articular cartilage using chondrocytes from articular cartilage have had little success, mainly because of improper
On the other hand, the CD271+CD73+ ectomesenchymal cells expressing
When and how the decision is made during chondrogenesis to generate growth plate‐like chondrocytes destined to form bone, or articular or nasal/auricular chondrocytes destined to form permanent cartilage are not yet fully understood. Research is undergoing to figure out the way to generate nasal chondrocytes from hPSC‐neural crest‐derived ectomesenchymal cells, which we believe will lead to answers to some of the questions.
4.2. Embryonic joint progenitor cells; articular chondroprogenitors or instructor cells for permanent cartilage formation
During embryogenesis, joint cartilage formation is initiated by the GDF5+ joint progenitor cells, which are distinct from the “general (SOX9+)” chondroprogenitors that can give rise to growth plate chondrocytes . Lineage tracing experiments demonstrate that the GDF5+ mesenchymal cells accumulate as a band within a cartilage anlage, constituting the interzone, or at the edges of the independently formed cartilage anlagen during joint (e.g., hip) formation [192, 193]. The joint progenitor cells expressing Wnt9a, Erg, and collagen IIA eventually give rise to articular cartilage, ligament, synovial lining, and other joint tissues but contribute little if at all to the growth plate cartilage. Isolated joint progenitor cells are induced to form chondroprogenitors, albeit weakly compared with the standard chondrocytes, in response to GDF5, and forced expression of Wnt9a during chondrogenesis is inhibitory. However, conditional deletion of β‐catenin inhibited the genesis of lubricin‐expressing, flat superficial zone‐like cell layer on articular cartilage , suggesting that canonical WNT signaling may change the chondrogenic fate of the joint progenitor from mid‐deep zone chondrocytes to superficial zone chondrocytes.
These results points to the potential usefulness of the joint progenitor cells for the repair of articular cartilage. Since the joint progenitors are characterized mainly during embryogenesis , it is conceivable to aim to generate, isolate, and characterize joint progenitor cells from PSCs. There is an interesting observation suggesting that the suppression of hedgehog signaling might promote genesis of the GDF5+ cell during mESC differentiation . As noted, hedgehog signaling, when not linked to the Ihh–PTHrP feedback mechanism, induces osteogenic gene expression in chondrocytes and facilitates endochondral ossification, which also causes osteoarthritic cartilage degradation in adult articular cartilage . Unfortunately, simple suppression of hedgehog signaling does not promote the development of
4.3. Postnatal joint stem/progenitor cells?
Consistent with GDF5+ embryonic joint progenitor cells being involved in the formation of a superficial layer of articular cartilage , the superficial zone chondrocytes show a distinct pattern of expression of stem/progenitor cell markers (Notch1, Stro1, vascular cell adhesion‐1, side population) [196–199]. Within the articular cartilage, Notch1 and Stro1 are expressed exclusively in the superficial zone, although Stro1 expression is not specific to articular cartilage, since the growth plate also abunduntly expresses Stro1 , and Notch1+ cells derived from the superficial zone are enriched in the colony‐forming unit fibroblast (CFU‐F) activity that is dependent on active Notch signaling . Therefore, the superficial zone has been hypothesized to harbor stem/progenitor cell activity. However, it has not been demonstrated whether such stem/progenitor cells can behave as resident joint stem/progenitor cells and “regenerate or repair” articular cartilage more effectively than the conventional adult chondrogenic cells such as bone marrow MSCs.
Another area of potential interest with regard to joint stem/progenitor cells is the perichondrial groove of Ranvier (or the zone of Ranvier). The area is located at the periphery of growth plate and is enriched in proliferating cells. Studies by Karlsson et al.  on the knee of adult rabbits have proved the existence of different subpopulations of progenitor cells in articular cartilage and the perichondrial groove of Ranvier. As with the superficial zone chondrocytes of articular cartilage, the cells in the groove of Ranvier express markers associated with stem cells and their niche (e.g., Stro1 and Jagged1), whereas cells in the growth plate directly adjacent to it do not express many of such markers. Mice lacking Tgfbr2 signaling in developing limb bud mesenchyme fail to form interzone, resulting in the absence of interphalangeal joints  as well as tendons and ligaments . The Tgfbr2‐expressing cells are in fact first detected at embryonic day (E) 13.5 within the interphalangeal joint interzone. Interestingly, by E16.5, the Tgfbr2‐expressing cells are enriched in the perichondrial groove of Ranvier, in part of the superficial layer of articular cartilage, in the synovium, and in the tendon's enthuses, and they remain in the same area postnatally . Such Tgfbr2‐expressing cells are slow‐growing cells and exhibit stem cell traits in expressing joint progenitor markers.
The knowledge that joint progenitor‐like cells, whether in the superficial zone of articular cartilage or the perichondrial groove of Ranvier, are present in postnatal joints, combined with the establishment of molecular tools to detect them are likely to inform future studies on the biology of postnatal joint progenitor cells. Isolation and functional characterization of these joint progenitor candidates will not only open the possibilities for an alternative cell source for regenerative therapy for cartilage, but also for the discovery of specific small or large molecule therapeutics that facilitate the regenerative activity of endogenous joint progenitor cells. However, as far as cellular therapy or tissue engineering therapy is concerned, the limitations noted with autologous chondrocyte/MSC implantation will also apply to these stem/progenitor cell systems—they will require expansion to clinically relevant levels without the loss of their developmental potential.
5. Conclusion and future perspective
As expected, PSC‐derived chondrogenic progeny has shown the advantage over adult tissue‐derived MSCs of “expansion without loss of chondrogenic potential” (Figure 1). However, the permanent(‐like) cartilage‐forming activity residing in the uncultured paraxial mesodermal cells (Figure 3 PM) and the nasal cartilage‐forming activity that the ectomesenchymal cells should possess, appear not to be readily maintained during expansion, even under the defined CDM + FGF2 + SB431542 (FSB) condition. In this sense, there is still room for the improvement in the expansion culture system. Controlling hypertrophic differentiation during chondrogenesis from such expanded hPSC‐progeny by manipulating known signaling mechanisms or using novel cell populations may overcome the problem associated with expansion culture. To date, however, such an approach either has not been systematically examined, even for the widely used adult stem cells, or has failed to give positive results. Probably, mechanistic principles that direct the generation of articular or nasal cartilage from corresponding precursor cells need to be elucidated for the reproducible generation of permanent cartilage from MSCs or hPSC‐derived, expanded chondroprogenitors.
For many years, the major challenges to making the hPSC system practical for therapeutic purposes have been how to direct differentiation to the specific cell lineage of interest and isolate the target cell population. But the important next challenge will be to control the aging or maturation of cell types generated from hPSCs so as to increase their functionality when transplanted into host adult tissues. Lastly, we will discuss two challenges associated with the use of embryonic/fetal cells for adult tissue repair/regeneration, and their potential relevance to the repair of articular cartilage by hPSC‐derived progeny.
5.1. Overgrowth of embryonic/fetal progenitors
Human PSCs (both ESCs and iPSCs) are teratoma‐forming cells by definition. Therefore, contamination of “undifferentiated” teratoma‐forming cells in the population of “differentiated, functional” progeny has been the major concern with the use of hPSC‐derived progenitor cells for therapeutic purposes. Even though the standard hPSCs are what is called “primed” PSCs, that is, they show less ability to self‐renew as a single cell than mPSC‐like “naïve” PSCs, many strategies have been devised to remove undifferentiated hPSCs. The use of iPSCs is associated with the additional concern that they are generated by transduction of adult tissue cells with a set of reprogramming genes that include
High proliferative properties of embryonic/fetal cell types that are the main cell types derived
5.2. Development of “adult”‐type cells
As noted, the process of differentiation of PSCs in culture can mimic many aspects of early embryogenesis. However, the terminally differentiated, functional cell types derived from PSCs often represent the embryonic counterpart of the adult cell type of interest, suggesting that the
In addition, cardiac muscle cells and their precursors (i.e., cardiac stem/progenitor cells) have been developed from hPSCs [225, 226]. The integration of such cells into rodent heart muscle improved cardiac functions in rodent models of myocardial infarction [227–230]. However, as is the case for erythrocytes, embryonic and adult cardiomyocytes differ in the composition of their major functional protein, the myosin heavy chain (MHC). For example, in the mouse, the slow MHC (MHCβ or MYH6) is predominantly expressed in the ventricle of embryonic hearts, while fast MHC (MHCα or MYH7) is dominant in the adult heart. In humans, this switch in ventricular MHCs is less pronounced, and MHCβ persists in the adult ventricle. In addition, the spontaneous beating phenotype of the hPSC‐derived cardiomyocytes resembles fetal but not adult cardiomyocytes, and the majority of the hPSC‐derived cardiomyocytes fail to fully recapitulate the electrophysiological function of adult ventricular cardiomyocytes [225, 231]. The most critical limitation is their deficiency in IK1, the potassium inwardly rectifying channel. Since the hPSC‐derived, spontaneously beating, cardiomyocyte clusters can transiently serve as a pacemaker in a pig heart upon transplantation , their functional differences from adult cardiomyocytes may increase the risk of cardiac arrhythmias upon transplantation. Therefore, similar to the case of HSCs, a robust, reproducible method that generates from hPSCs cardiac stem/progenitor cells capable of giving rise to mature adult cardiomyocytes is needed if hPSCs are to be used for the repair of damaged heart muscle.
Where cartilage is concerned, no adult–fetal differences in the biochemical property of joint chondrocytes have been elucidated. Differences have only been observed in the cellularity and physical properties of child and adult articular cartilage. Since there appears to be no active replenishing of articular chondrocytes or spontaneous regeneration of damaged articular or meniscus cartilage in adult humans and large animals, we hypothesize that embryonic/fetal cells are likely to perform better in cartilage repair than adult cells.
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