Induction of E-cadherin+ human amniotic fluid cell differentiation into oocyte-like cells via culture in medium supplemented with follicular fluid.
Pluripotent human amniotic fluid cells (HuAFCs) can differentiate into various types of somatic cell in vitro. However, their differentiation into oocyte-like cells has never been described to the best of our knowledge. In the present study, differentiation of E-cadherin+ and E-cadherin- HuAFC sub-populations into oocyte-like cells was induced via culture in medium containing bovine follicular fluid and β-mercaptoethanol. The E-cadherin+ HuAFCs expressed DAZL highly. Post-induction, cells with an oocyte-like phenotype were found among the E-cadherin+ HuAFCs, expressing markers specific to germ cells and oocytes (VASA, ZP3 and GDF9) and meiosis (DMC1 and SCP3). When specific small interfering RNA (siRNA) was used to suppress E-cadherin in the E-cadherin+ HuAFCs, the levels of DAZL expression were reduced. Post-induction, the morphology of the siRNA‑E‑cadherin HuAFCs was poorer and the expression levels of germ cell-specific markers were lower compared with those of the siRNA-mock HuAFCs. Therefore, E-cadherin+ HuAFCs could be more easily induced to differentiate into oocyte-like cells by bovine follicular fluid and β-mercaptoethanol. In addition, the E-cadherin+ HuAFCs exhibited potential characteristics of DAZL protein expression, and thus it was conjectured that bovine follicular fluid acts on DAZL protein and promotes E-cadherin+ HuAFC differentiation into oocyte-like cells.
Shanghai Geriatric Institute of Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200031, P.R. China.
Te
T
Liu
School of Life Science and Technology, Tongji University, Shanghai 200092, P.R. China.
Yongyi
Y
Huang
College of Life Science, Henan Normal University, Xinxiang 453007, P.R. China.
Yanzhen
Y
Bu
Department of Oral and Craniofacial Science, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China.
Yanhui
Y
Zhao
Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, P.R. China.
Gang
G
Zou
School of Life Science and Technology, Tongji University, Shanghai 200092, P.R. China.
Zhixue
Z
Liu
D000293:Adolescent; D000653:Amniotic Fluid / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D015820:Cadherins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry / Q000378:metabolism; D055429:Growth Differentiation Factor 9 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008623:Mercaptoethanol / Q000494:pharmacology; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D011247:Pregnancy; D034741:RNA, Small Interfering / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D055815:Young Adult; D000072216:Zona Pellucida Glycoproteins
D015820:Cadherins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D004527:Egg Proteins; C525897:GDF9 protein, human; D055429:Growth Differentiation Factor 9; D008562:Membrane Glycoproteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface; C490060:SYCP3 protein, human; C000606517:ZP3 protein, human; D000072216:Zona Pellucida Glycoproteins; D008623:Mercaptoethanol; C498288:DDX4 protein, human; C074556:DMC1 protein, human; D053487:DEAD-box RNA Helicases