TY - JOUR
T1 - Sorting and Manipulation of Human PGC-LC Using PDPN and Hanging Drop Cultures
AU - Arkoun, Brahim
AU - Moison, Pauline
AU - Guerquin, Marie Justine
AU - Messiaen, Sébastien
AU - Moison, Delphine
AU - Tourpin, Sophie
AU - Monville, Christelle
AU - Livera, Gabriel
N1 - Publisher Copyright:
© 2022 by the authors.
PY - 2022/12/1
Y1 - 2022/12/1
N2 - The generation of oocytes from induced pluripotent stem cells (iPSCs) was proven efficient with mouse cells. However, no human iPSCs have yet been reported to generate cells able to complete oogenesis. Additionally, efficient sorting of human Primordial Germ Cell-like Cells (hPGC-LCs) without genomic integration of fluorescent reporter for their downstream manipulation is still lacking. Here, we aimed to develop a model that allows human germ cell differentiation in vitro in order to study the developing human germline. The hPGC-LCs specified from two iPS cell lines were sorted and manipulated using the PDPN surface marker without genetic modification. hPGC-LCs obtained remain arrested at early stages of maturation and no further differentiation nor meiotic onset occurred when these were cultured with human or mouse fetal ovarian somatic cells. However, when cultured independently of somatic ovarian cells, using BMP4 and the hanging drop-transferred EBs system, early hPGC-LCs further differentiate efficiently and express late PGC (DDX4) and meiotic gene markers, although no SYCP3 protein was detected. Altogether, we characterized a tool to sort hPGC-LCs and an efficient in vitro differentiation system to obtain pre-meiotic germ cell-like cells without using a gonadal niche.
AB - The generation of oocytes from induced pluripotent stem cells (iPSCs) was proven efficient with mouse cells. However, no human iPSCs have yet been reported to generate cells able to complete oogenesis. Additionally, efficient sorting of human Primordial Germ Cell-like Cells (hPGC-LCs) without genomic integration of fluorescent reporter for their downstream manipulation is still lacking. Here, we aimed to develop a model that allows human germ cell differentiation in vitro in order to study the developing human germline. The hPGC-LCs specified from two iPS cell lines were sorted and manipulated using the PDPN surface marker without genetic modification. hPGC-LCs obtained remain arrested at early stages of maturation and no further differentiation nor meiotic onset occurred when these were cultured with human or mouse fetal ovarian somatic cells. However, when cultured independently of somatic ovarian cells, using BMP4 and the hanging drop-transferred EBs system, early hPGC-LCs further differentiate efficiently and express late PGC (DDX4) and meiotic gene markers, although no SYCP3 protein was detected. Altogether, we characterized a tool to sort hPGC-LCs and an efficient in vitro differentiation system to obtain pre-meiotic germ cell-like cells without using a gonadal niche.
KW - human germline
KW - Induced pluripotent stem cells
KW - meiotic commitment
KW - primordial germ cells
UR - http://www.scopus.com/inward/record.url?scp=85143651163&partnerID=8YFLogxK
U2 - 10.3390/cells11233832
DO - 10.3390/cells11233832
M3 - Article
C2 - 36497094
AN - SCOPUS:85143651163
SN - 2073-4409
VL - 11
JO - Cells
JF - Cells
IS - 23
M1 - 3832
ER -