胎盘贴壁细胞的生物学特性

Biological characteristics of placenta-derived adherent cells

目的：通过体外分离和培养胎盘贴壁细胞，以研究其形态、表型和分化特点．方法：采用Ⅳ型胶原酶消化人胎盘组织，获得单个核细胞，接种于100mL／LFBS低糖DMEM培养基中，贴壁后常规换液、传代，流式细胞仪检测其表面标志，绘制生长曲线．采用β-甘油磷酸钠、维生素C、地塞米松联合诱导，使胎盘贴壁细胞分化为成骨细胞，诱导后10d行茜素红染色；采用地塞米松、胰岛素诱导，使胎盘贴壁细胞分化为脂肪细胞，油红O染色鉴定．结果：培养7～14d后有少量贴壁细胞生长，逐渐呈成纤维细胞状，表达CD29，CD44和CD105，不表达CD34，CD45，CD19；在成骨诱导10d后细胞茜素红染色可见钙盐沉积，成脂肪诱导7d后油红O染色见脂肪滴形成．结论：通过不同方式诱导培养，胎盘贴壁细胞可分化为成骨细胞及脂肪细胞，提示胎盘贴壁细胞可能是新的干细胞来源．

AIM： To isolate and culture human placenta-derived adherent cells （hPDAC） in vitro and study their biological characteristics involving morphology, phenotype and differentiation. METHODS： Mononuclear cells were harvested after placenta tissue was digested by collagenase Ⅳ, seeded and cultured in low-glucose DMEM containing 100 mL/L FBS. Their surface markers were detected by flow cytometry; the growth curve was made; cells of 3 passages were induced to differentiate into osteoblasts with vitamin C, dexamethasone and sodium β-glycerophosphate, and the induced cells were observed in the change of matrix mineralization with alizarin red staining; cells of 3 passages were induced to differentiate into lipocytes with dexamethasone and insulin, and the induced cells were observed with oil red O staining. RESULTS ： By day 7 - 14, a few of adherent cells were observed and expanded the same as fibroblasts, and adherent cells were positive for CD29, CD44 and CD105 , but negative for CD34, CD45 , CD19. After 10-day induction, alizarin red staining showed the calcium mineralization and after 7 d, oil red O staining revealed the appearance of fat drops. CONCLUSION： The hPDAC was confirmed to have the osteogenic and adipogenic potentials, indicating that they could be regarded as an alternative source of stem cells.