title: Human amniotic fluid stem cells have a potential to recover ovarian function in mice with chemotherapy-induced sterility.
abstract: BACKGROUND
  Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine. The ability of hAFCs to differentiate into germ cell and oocyte-like cells has been previously documented. Herein we report the potential use of hAFCs to help restore follicles in clinical condition involving premature ovarian failure.
  
  
  RESULTS
  Human amniotic fluid was obtained via amniocentesis, yielding a subpopulation of cloned hAFCs that was able to form embryoid bodies (EBs) and differentiate into three embryonic germ layers. Moreover, culture of EBs in medium containing human follicular fluid (HFF) or a germ cell maturation factor cocktail (FAC), expressed germ cells markers such as BLIMP1, STELLA, DAZL, VASA, STRA8, SCP3, SCP1, and GDF9. Furthermore, one cell line was grown from clone cells transfected with lentivirus-GFP and displaying morphological characteristics of mesenchymal cells, had the ability to restore ovarian morphology following cell injection into the ovaries of mice sterilized by intraperitoneal injection of cyclophosphamide and busulphan. Restored ovaries displayed many follicle-enclosed oocytes at all stages of development, but no oocytes or follicles were observed in sterilized mice whose ovaries had been injected with medium only (control). Notably, identification of GFP-labeled cells and immunostaining with anti-human antigen-specific antibodies demonstrated that grafted hAFCs survived and differentiated into granulosa cells which directed oocyte maturation. Furthermore, labeling of ovarian tissue for anti-Müllerian hormone expression, a functional marker of folliculogenesis, was strong in hAFCs-transplanted ovaries but inexistent in negative controls.
  
  
  CONCLUSIONS
  These findings highlight the possibility of using human amniotic fluid-derived stem cells in regenerative medicine, in particular in the area of reproductive health.
authors.affiliation: The International Peace Maternity and Child Health Hospital, School of medicine, Shanghai Jiaotong University, Shanghai 200030, China. laidongmei@hotmail.com.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Lai
authors.forename: Fangyuan
authors.initials: F
authors.lastname: Wang
authors.forename: Yifei
authors.initials: Y
authors.lastname: Chen
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Yanlin
authors.initials: Y
authors.lastname: Wang
authors.forename: Weiwei
authors.initials: W
authors.lastname: Cheng
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology* / Q000378:metabolism*; D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000009:adverse effects*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003520:Cyclophosphamide / Q000008:administration & dosage / Q000009:adverse effects*; D004195:Disease Models, Animal; D058732:Embryoid Bodies / Q000166:cytology; D005260:Female; D006801:Humans; D007247:Infertility, Female / Q000139:chemically induced / Q000628:therapy*; D059785:Karyotype; D059630:Mesenchymal Stem Cells / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D010050:Ovarian Function Tests; D010053:Ovary / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000502:physiology* / Q000637:transplantation; D011247:Pregnancy; D033581:Stem Cell Transplantation
chemical_list: D000970:Antineoplastic Agents; D003520:Cyclophosphamide