title: cGMP-Compliant Expansion of Human iPSC Cultures as Adherent Monolayers.
abstract: Therapeutic uses of cells differentiated from human pluripotent stem cells (hPSCs), either embryonic stem (ES) cells or induced pluripotent stem cells (iPSCs), are now being tested in clinical trials, and it is likely that this will lead to increased commercial interest in the clinical translation of promising hPSC research. Recent technical advances in the use of defined media and culture substrates have significantly improved both the simplicity and predictability of growing hPSCs, allowing a much more straightforward application of current good manufacturing practices (cGMP) to the culture of these cells. In addition, the adoption of cGMP-compliant techniques in research environments will both improve the replication of results and make the transition of promising investigations to the commercial sector significantly less cumbersome. However, passaging methods for hPSCs are inherently unpredictable and rely on operator experience and expertise. This is problematic for the cell manufacturing process where operator time and process predictability are often determining cost drivers. We have adopted a human iPSC system using defined media and a recombinant substrate that employs cell dissociation with a hypertonic citrate solution which eliminates variability during hPSC cell expansion and provides a simple cGMP-compliant technique for hiPSC cultivation that is appropriate in both research and commercial applications.
authors: Department of Neurosurgery, 420 Delaware Street, MMC 96, Minneapolis, 55455, USA.
authors: Ann M
authors: AM
authors: Parr
authors: Stem Cell Institute, University of Minnesota, 2001 6th Street SE, Minneapolis, MN, 55455, USA.
authors: Patrick J
authors: PJ
authors: Walsh
authors: Stem Cell Institute, University of Minnesota, 2001 6th Street SE, Minneapolis, MN, 55455, USA.
authors: Vincent
authors: V
authors: Truong
authors: Stem Cell Institute, University of Minnesota, 2001 6th Street SE, Minneapolis, MN, 55455, USA. dutto015@umn.edu.
authors: James R
authors: JR
authors: Dutton
mesh_terms: D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods; D002614:Chelating Agents / Q000494:pharmacology; D002951:Citrates / Q000494:pharmacology; D003470:Culture Media; D005347:Fibroblasts / Q000166:cytology; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D006982:Hypertonic Solutions / Q000494:pharmacology; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D000077559:Sodium Citrate; D019096:Vitronectin / Q000494:pharmacology
chemical_list: D002614:Chelating Agents; D002951:Citrates; D003470:Culture Media; D006982:Hypertonic Solutions; D011994:Recombinant Proteins; D019096:Vitronectin; D000077559:Sodium Citrate
keywords: Cell manufacturing; Pluripotent stem cells; iPS cell culture