Human somatic cells subjected to genetic induction with six germ line-related factors display meiotic germ cell-like features.
The in vitro derivation of human germ cells has attracted interest in the last years, but their direct conversion from human somatic cells has not yet been reported. Here we tested the ability of human male somatic cells to directly convert into a meiotic germ cell-like phenotype by inducing them with a combination of selected key germ cell developmental factors. We started with a pool of 12 candidates that were reduced to 6, demonstrating that ectopic expression of the germ line-related genes PRDM1, PRDM14, LIN28A, DAZL, VASA and SYCP3 induced direct conversion of somatic cells (hFSK (46, XY), and hMSC (46, XY)) into a germ cell-like phenotype in vitro. Induced germ cell-like cells showed a marked switch in their transcriptomic profile and expressed several post-meiotic germ line related markers, showed meiotic progression, evidence of epigenetic reprogramming, and approximately 1% were able to complete meiosis as demonstrated by their haploid status and the expression of several post-meiotic markers. Furthermore, xenotransplantation assays demonstrated that a subset of induced cells properly colonize the spermatogonial niche. Knowledge obtained from this work can be used to create in vitro models to study gamete-related diseases in humans.
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.|Fundación Instituto de Investigación Sanitaria La Fe, Valencia, 46026, Spain.
Jose V
JV
Medrano
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.
Ana M
AM
Martínez-Arroyo
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.
Jose M
JM
Míguez
Igenomix S.L., Paterna, 46980, Spain.
Inmaculada
I
Moreno
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.
Sebastián
S
Martínez
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.
Alicia
A
Quiñonero
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.
Patricia
P
Díaz-Gimeno
Valencia Node of the Spanish Stem Cell Bank, Prince Felipe Research Centre (CIPF) Valencia, 46012, Spain.
Ana I
AI
Marqués-Marí
Fundación Instituto de Investigación Sanitaria La Fe, Valencia, 46026, Spain.
Antonio
A
Pellicer
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.
Jose
J
Remohí
Fundación Instituto Valenciano de Infertilidad (FIVI), INCLIVA, Department of Pediatrics, Obstetrics and Gynecology, Valencia University, Valencia, 46015, Spain.|Igenomix S.L., Paterna, 46980, Spain.|Department of Obstetrics and Gynecology, Stanford University, Stanford, CA 94305, USA.
Carlos
C
Simón
D000818:Animals; D002066:Busulfan / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004171:Diploidy; D052816:Foreskin; D005854:Germ Cells / Q000166:cytology / Q000502:physiology*; D006238:Haploidy; D006801:Humans; D008297:Male; D008540:Meiosis; D008819:Mice, Nude; D013093:Spermatogonia / Q000166:cytology / Q000502:physiology*; D015533:Transcriptional Activation / Q000502:physiology*; D059467:Transcriptome
D002066:Busulfan