title: Direct differentiation of human pluripotent stem cells into haploid spermatogenic cells.
abstract: Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) have been shown to differentiate into primordial germ cells (PGCs) but not into spermatogonia, haploid spermatocytes, or spermatids. Here, we show that hESCs and hiPSCs differentiate directly into advanced male germ cell lineages, including postmeiotic, spermatid-like cells, in vitro without genetic manipulation. Furthermore, our procedure mirrors spermatogenesis in vivo by differentiating PSCs into UTF1-, PLZF-, and CDH1-positive spermatogonia-like cells; HIWI- and HILI-positive spermatocyte-like cells; and haploid cells expressing acrosin, transition protein 1, and protamine 1 (proteins that are uniquely found in spermatids and/or sperm). These spermatids show uniparental genomic imprints similar to those of human sperm on two loci: H19 and IGF2. These results demonstrate that male PSCs have the ability to differentiate directly into advanced germ cell lineages and may represent a novel strategy for studying spermatogenesis in vitro.
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Pittsburgh School of Medicine, Pittsburgh, PA 15108, USA.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Easley
authors.forename: Bart T
authors.initials: BT
authors.lastname: Phillips
authors.forename: Megan M
authors.initials: MM
authors.lastname: McGuire
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Barringer
authors.forename: Hanna
authors.initials: H
authors.lastname: Valli
authors.forename: Brian P
authors.initials: BP
authors.lastname: Hermann
authors.forename: Calvin R
authors.initials: CR
authors.lastname: Simerly
authors.forename: Aleksander
authors.initials: A
authors.lastname: Rajkovic
authors.forename: Toshio
authors.initials: T
authors.lastname: Miki
authors.forename: Kyle E
authors.initials: KE
authors.lastname: Orwig
authors.forename: Gerald P
authors.initials: GP
authors.lastname: Schatten
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D006238:Haploidy; D006801:Humans; D008297:Male; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D013087:Spermatids / Q000166:cytology / Q000378:metabolism*; D013090:Spermatocytes / Q000166:cytology / Q000378:metabolism*; D013091:Spermatogenesis / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D014157:Transcription Factors