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目的按照Kobayashi方法从羊膜间充质细胞(AMCs)的侧群细胞(SP)中分离羊膜干细胞并予以细胞学特性鉴定。方法按文献法取剖腹产手术胎盘,用色素染色法和流式细胞分选技术从AMCs中分离羊膜干细胞进行原代培养,用PI染色DNA分析细胞周期,软琼脂培养法观察细胞体外集落,并与人肝癌细胞系HepG2比较。结果1.从AMCs中成功分离的(羊膜干细胞侧群细胞)AMCs-SP约占总AMCs的0.2%;2.细胞周期结果为AMC-SP细胞增殖缓慢或处于静止期;3.体外培养时细胞群体培增数达40以上,在软琼脂中AMC-SP不形成集落,与人肝癌细胞系HepG2可形成较大集落形成反差。结论AMC-SP具有细胞来源充足、稳定性好、安全性高、无伦理学争议,异体移植无急性免疫排外反应,无致瘤性的优点,可做为再生医学及急性损伤期疾病治疗的重要的人细胞来源应用于临床。
OBJECTIVE To isolate amniotic stem cells from the lateral mesenchymal cells (SPs) of amniotic mesenchymal cells (AMCs) according to the Kobayashi method and identify their cytological characteristics. Methods According to the literature method, cesarean section placenta was taken, and amniotic membrane stem cells were isolated from AMCs by dye staining and flow cytometry. Primary cultured cells were stained with PI staining DNA. Cell colony was observed by soft agar culture. Comparison of human hepatoma cell line HepG2. The results showed that AMCs-SP cells isolated from AMCs successfully accounted for about 0.2% of the total AMCs. 2. The cell cycle results showed that the proliferation of AMC-SP cells was slow or in quiescence. 3. The cells cultured in vitro The population of cultivars increased by more than 40, AMC-SP did not form colonies in soft agar and formed colony contrast with human hepatoma cell line HepG2. Conclusion AMC-SP has the advantages of sufficient source of cells, good stability, high safety, no ethical controversy, no acute rejection after xenotransplantation and no tumorigenicity. It can be used as an important medicine for regenerative medicine and acute injury treatment The source of human cells used in clinical.