Una Chen

By Una Chen

Together with her colleagues, the author has worked extensively on the differentiation potential of mouse embryonic stem cells (ESCs) to blood islands containing embryoid bodies (EBs) in culture. Such EBs contain hematopoietic stem- precursor cells (HSCs) committed to lymphoid, myeloid lineages and cells possessing Natural Killer (NK) cell marker. Further differentiation of such EBs to mature B-lymphoid cells can be demonstrated in a second stage of tissue culture, by co-culturing separated EBs with a mitogen, Lipopolysaccharide (LPS). The demonstration of differentiation of such cells to mature T- and B-lymphoid cells was performed in vivo, i.e., by implanting EBs subcutaneously in nude mice. The differentiation of ESCs to EBs needed to be scaled up to obtain enough HSC-containing EBs for a second stage of differentiation, in vitro and in vivo. Selection of HSC-containing EBs must be performed manually under a microscope outside the tissue culture incubator. For research purposes, the cell culture medium contains antibiotics to inhibit the growth of bacteria. However for clinical application, the medium should be free of antibiotics. A conventional research laboratory environment is not sufficient for such an application. A GMP (Good Manufacturing Production) facility with clean air became necessary. Commercial clean air facilities used in the operation room and/or by industry were over budget. The attention of the author has shifted to developing an affordable GMP facility for clinical research in academia. These efforts were successful and the price for such a GMP facility is 10% of a comparable industrial model. After that experience, “a GMP facility for cell therapy on wheels” was further developed. This chapter will describe and share with academic colleagues our experience of constructing such affordable GMP facilities from scratch, starting with the purchase of high-efficiency particulate arrestance (HEPA) filters for the clean air facility.

Part of the book: Pluripotent Stem Cells