RT Journal Article
SR Electronic
T1 Efficient Human Germ Cell Specification from Stem Cells via Combinatorial Expression of Transcription Factors
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.07.11.499564
DO 10.1101/2022.07.11.499564
A1 Christian Kramme
A1 Merrick Pierson Smela
A1 Bennett Wolf
A1 Patrick R. Fortuna
A1 Garyk Brixi
A1 Kalyan Palepu
A1 Edward Dong
A1 Jessica Adams
A1 Suhaas Bhat
A1 Sabrina Koseki
A1 Emma Tysinger
A1 Teodora Stan
A1 Richie E. Kohman
A1 Songlei Liu
A1 Mutsumi Kobayashi
A1 Toshi Shioda
A1 George M. Church
A1 Pranam Chatterjee
YR 2022
UL http://biorxiv.org/content/early/2022/07/12/2022.07.11.499564.abstract
AB Germ cells are the vehicle of human reproduction, arising early in embryonic development and developing throughout adult life until menopause onset in women. Primordial germ cells are the common precursors of germline cells in both sexes, undergoing sexual specification into oogonia or gonocytes which further develop into oocytes or spermatocytes during development. Methods for recapitulation of primordial germ cell and oogonia formation have been developed extensively in recent decades, but fundamental technical limitations in their methodologies, throughput, and yield limit their utilization. Recently, transcription factor (TF)-based methods for human primordial germ cell-like cell (hPGCLC) formation, mouse meiotic entry, and mouse oocyte maturation have demonstrated the feasibility of gene overexpression screening in identifying potent regulators of germ cell development. Here we screened 47 folliculogenesis-regulating TFs for their role in hPGCLC and oogonia formation, identifying DLX5, HHEX, and FIGLA whose individual overexpression enhances hPGCLC formation from hiPSCs. Additionally, we identify a set of three TFs, ZNF281, LHX8, and SOHLH1, whose combinatorial overexpression drives direct oogonia-like formation from hiPSCs in a four-day, feeder-free monolayer culture condition with additional feeder-free culture capabilities post-isolation. We characterize these TF-based germ cells via gene and protein expression analyses, and demonstrate their broad similarity to in vivo germ cells. Together, these results identify novel regulators of human germ cell development and establish new TF-based tools for human in vitro oogenesis research.Competing Interest StatementP.C., C.K., M.P.S., and G.C. are listed as inventors for U.S. Provisional Application No. 63/326,656, entitled: “Methods and Compositions for Producing Primordial Germ Cell-Like Cells,” and U.S. Provisional Application No. 63/326,607, entitled: “Methods and Compositions for Producing Oogonia-Like Cells.” P.C. is a co-founder and scientific advisor to Gameto, Inc. C.K. is the Chief Scientific Officer of Gameto, Inc. G.M.C. serves on the scientific advisory board of Gameto, Inc., Colossal Biosciences, and GCTx.