title: Platform for induction and maintenance of transgene-free hiPSCs resembling ground state pluripotent stem cells.
abstract: Cell banking, disease modeling, and cell therapy applications have placed increasing demands on hiPSC technology. Specifically, the high-throughput derivation of footprint-free hiPSCs and their expansion in systems that allow scaled production remains technically challenging. Here, we describe a platform for the rapid, parallel generation, selection, and expansion of hiPSCs using small molecule pathway inhibitors in stage-specific media compositions. The platform supported efficient and expedited episomal reprogramming using just OCT4/SOX2/SV40LT combination (0.5%-4.0%, between days 12 and 16) in a completely feeder-free environment. The resulting hiPSCs are transgene-free, readily cultured, and expanded as single cells while maintaining a homogeneous and genomically stable pluripotent population. hiPSCs generated or maintained in the media compositions described exhibit properties associated with the ground state of pluripotency. The simplicity and robustness of the system allow for the high-throughput generation and rapid expansion of a uniform hiPSC product that is applicable to industrial and clinical-grade use.
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Bahram
authors.initials: B
authors.lastname: Valamehr
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Megan
authors.initials: M
authors.lastname: Robinson
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Ramzey
authors.initials: R
authors.lastname: Abujarour
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Betsy
authors.initials: B
authors.lastname: Rezner
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Florin
authors.initials: F
authors.lastname: Vranceanu
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Thuy
authors.initials: T
authors.lastname: Le
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Amanda
authors.initials: A
authors.lastname: Medcalf
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Tom Tong
authors.initials: TT
authors.lastname: Lee
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Fitch
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: David
authors.initials: D
authors.lastname: Robbins
authors.affiliation: Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Flynn
mesh_terms: D059786:Abnormal Karyotype; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D002869:Chromosome Aberrations; D016000:Cluster Analysis; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005347:Fibroblasts; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D042822:Genomic Instability; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D019076:Transgenes
chemical_list: D014157:Transcription Factors