title: Derivation and propagation of hESC under a therapeutic environment.
abstract: The pluripotent nature of human embryonic stem cells (hESC) makes them very attractive as a source of various cell types that could be used therapeutically in regenerative medicine. However, eliminating all sources of contamination, animal-derived or human cell-derived, during hESC derivation and propagation is necessary before hESC derivatives can be used clinically. Although there is continuing progress toward this goal, none of the methods to date to produce hESC lines under good manufacturing practices (GMP) has been published. The long-term success for GMP compliance depends critically on maintaining and implementing a stringent quality control system which is also dictated by the regulatory authorities in different countries. In this unit, an approach is described based upon the experience of this author and others towards achieving clinical-grade hESC lines systematically involving all the steps from start to finish under GMP environment. This unit provides a basic layout for GMP set up to achieve quality controls, a step-by-step guide to producing new hESC lines under defined conditions, and standard operating procedures used to achieve this outcome.
authors.affiliation: Diabetes Transplant Unit, The Prince of Wales Hospital and The University of New South Wales, New South Wales, Australia.
authors.forename: Kuldip S
authors.initials: KS
authors.lastname: Sidhu
authors.forename: Sarah
authors.initials: S
authors.lastname: Walke
authors.forename: Bernard E
authors.initials: BE
authors.lastname: Tuch
mesh_terms: D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D017364:Collagenases / Q000494:pharmacology; D016895:Culture Media, Serum-Free; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D010450:Endopeptidases / Q000494:pharmacology; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D005452:Fluoresceins / Q000378:metabolism; D005615:Freezing; D006801:Humans; D007202:Indicators and Reagents; D011419:Propidium / Q000378:metabolism; D013997:Time Factors
chemical_list: D016895:Culture Media, Serum-Free; D005452:Fluoresceins; D007202:Indicators and Reagents; D011419:Propidium; D010450:Endopeptidases; D017364:Collagenases; C023139:dispase