人腹膜中间充质干细胞生物学特性的初步研究

目的探讨人腹膜中是否含有间充质干细胞(Mesenchymal stem cells,MSC),并研究其生物学特性。方法腹膜取自接受胃大部切除及腹腔淋巴结清扫术的胃癌患者。将腹膜剪碎,Ⅰ型胶原酶消化后以低密度接种,2周后计算细胞群中成纤维细胞集落形成单位(Colony-forming unit-fibroblast,CUF-F)数量;流式细胞分析贴壁细胞表型;定向诱导2周后,碱性磷酸酶和油红O染色,观察细胞体外成骨和成脂肪能力。结果贴壁的细胞呈成纤维细胞样,消化的腹膜细胞群中,CFU-F比例平均为0.574‰(0.26‰～0.84‰)。流式分析显示,细胞均一表达CD29、CD44和CD73,不表达CD31、CD34和CD45。成骨细胞定向诱导后,细胞呈现碱性磷酸酶活性;成脂诱导后细胞内出现脂肪滴,油红O染色阳性。结论人腹膜中富含MSC,可作为一个含有生物支架的干细胞来源。

The Biological Properties of Mesenchymal Stem Cells Derived from Human Peritoneum

Objective To observe the existence of mesenchymal stem cells（MSCs） in the human peritoneum and investigate their biological characteristics.Methods The peritoneum was harvested from five patients with gastric cancer.The peritoneum was cut into pieces and digested by collagenase type Ⅰ.The harvested cells were seeded at a low density and the number of colony-forming unit-fibroblast was counted after culturing 14 days.The phenotypic features of the adherent cells were analyzed by flow cytometric techniques.Their osteogenesis and adipogenesis in vitro were evaluated by alkaline phosphatase and Oil Red O staining after 2 weeks of directional induction.Results The adherent cells were fibroblast-like in morphology,and the proportion of CFU-F in the digested cells was 0.574‰（0.26‰-0.84‰）.The expressions of CD29,CD44 and CD73 were positive while the expressions of CD31,CD34 and CD45 were negative.The cells were positive for alkaline phosphatase after osteogenic induction.Meanwhile,intracellular lipid droplets appeared after adipogenic induction and was shown positive by Oil-Red O staining.Conclusion MSCs are rich in human peritoneum and they might be served as a source for stem cells with natural scaffolds.