In vitro female germline potential of human umbilical cord-derived matrix stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Bone marrow mesenchymal stem cells(BM-MSCs)have been shown to possess the potential to differentiate into oocytes.However,immune rejection and a limited number of donors of BM-MSCs constrain the applications of BM-MSCs.Several studies have demonstrated that human umbilical cord matrix stem cells(UC-MSCs)also have an intrinsic ability to differentiate into oocyte-like cells in vitro.OBJECTIVE:To establish the method for UC-MSCs culture and to investigate the in vitro differentiation potential of UC-MSCs towards germ cells.METHODS:Umbilical cord from full-term normal deliveries was obtained in sterile condition.Collagenase I-digested cells were cultured in DMEM.The immunophenotype of cells was determined by flow cytometry.Lipoblasts,osteoblasts and chondroblasts were induced in different condition cultures.The expression of germ cells specific marker in UC-MSCs was determined by reverse transcdption-polymerase chain reaction.Follicular fluid was employed to induce UC-MSCs differentiation into germ cells.RESULTS AND CONCLUSION:Spindle-like umbilical cord cells were shown and cells in culture were extended to more than 10passages.BM-MSCs-like immunophenotypes were shownCD29,CD44,CD73(SH3),CD90 and CD105(SH2)were positive;SSEA-4 was weakly positive;CD31,CD34,CD45 and HLA-DR were negative.After UC-MSCs were induced in different condition cultures,lipid droplet-,bone tubercle-,and cartilage tubercle-like structures emerged and the mRNA expressions of specific gene of fat,bone and cartilage were observed.Germ cells markers,OCT4,Stella,Ifitm3,were expressed in UC-MSCs.After induced by 5%,10% or 20% follicular fluid,cells aggregated and oocyte-like structures were observed.Human UC-MSCs could be cultured and amplificated in vitro.UC-MSCs showed immunophenotypes similar to BM-MSCs.UC-MSCs had the potential to differentiate into lipoblasts,osteoblasts,and chondroblasts.Oocyte-like structure was induced in vitro from UC-MSCs with germ cells specific marker.These findings suggest that UC-NSCs have the potential to differentiate into germ cells.