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羊水干细胞是一种存在于胎儿羊水中Oct4阳性的多能性干细胞。该细胞具有增殖能力强、分化潜能高和免疫原性低等特点,在生物学和医学研究上具有广泛的应用前景。本实验从不同胎龄的猪胎儿标本中分离猪羊水干细胞,并通过诱导其向脂肪细胞分化来检测其多向分化潜能。结果表明,从胎龄为30～70d的猪胎儿羊水中分离出的羊水干细胞贴壁率高,增殖能力强,活性好,并建立了两株猪羊水干细胞系。经流式细胞仪和RT-PCR检测,猪羊水干细胞表达CD117、CD44、CD166和CD90表面抗原,不表达CD45、CD54和CD34;稳定表达Oct4、Nanog、C-myc和HLA-abc干细胞特异标志基因;转染报告基因载体pOct4-EGFP和pNanog-EGFP后表达绿色荧光;此外,猪羊水干细胞还能够在体外诱导分化为脂肪细胞。研究结果为大动物羊水干细胞的研究提供了可借鉴的材料。
Amniotic fluid stem cells are Oct4-positive pluripotent stem cells present in fetal amniotic fluid. The cell has the characteristics of strong proliferation, high differentiation potential and low immunogenicity, and has broad application prospects in biology and medical research. In this study, porcine amniotic fluid stem cells were isolated from porcine fetal specimens of different gestational ages and their multipotential differentiation potential was detected by inducing them to differentiate into adipocytes. The results showed that amniotic fluid stem cells isolated from fetal amniotic fluid of gestational age of 30 ~ 70d had high adherent rate, strong proliferative capacity and good activity, and established two strains of porcine amniotic fluid stem cells. The results of flow cytometry and RT-PCR showed that porcine amniotic fluid stem cells expressed CD117, CD44, CD166 and CD90 surface antigens but did not express CD45, CD54 and CD34. Stable expression of Oct4, Nanog, C-myc and HLA-abc stem cell specific marker genes ; Transfection of reporter gene vectors pOct4-EGFP and pNanog-EGFP expression of green fluorescence; In addition, porcine amniotic fluid stem cells can also be induced in vitro differentiation of adipocytes. The results provide a reference for the research of amniotic fluid stem cells in large animals.