脂肪干细胞体外分离培养鉴定及免疫学性质

Human adipose-derived stem cells: in vitro isolation, culture, identification and immunological properties

背景:脂肪干细胞由于具备易于获取等特点,在组织工程中比骨髓间充质干细胞具有更加广阔的应用前景,并因此成为当今的研究热点之一。目的:观察脂肪干细胞的生长规律,鉴定其干细胞特性和免疫学性质。方法:自整形外科行抽脂的3名健康志愿者身上获取皮下脂肪组织及外周血,志愿者年龄20-30岁,均为女性。用Ⅰ型胶原酶消化脂肪组织获得脂肪干细胞,在体外原代培养及传代培养,观察其体外增殖能力等生物学特性;通过流式细胞术检测人脂肪干细胞分化群表面抗原和主要组织相容性抗原的表达。并将异体脂肪干细胞和淋巴细胞共培养,观察脂肪干细胞是否可以刺激后者增殖。结果与结论:所获脂肪干细胞具有很强的体外增殖能力,其生长曲线呈"S"形;第3代脂肪干细胞表面CD29、CD44、CD90以及主要组织相容性抗原Ⅰ呈阳性表达,CD31、CD45、CD34及主要组织相容性抗原Ⅱ呈阴性表达;未发现异体脂肪干细胞刺激淋巴细胞增殖。提示所获细胞确为脂肪干细胞,且具有较弱的排斥反应,有可能成为细胞治疗或组织工程的同种异体细胞来源。

BACKGROUND： Adipose-derived stem cells have more broad application prospects than bone marrow mesenchymal stem cells in tissue engineering because of the characteristics such as easy to obtain, and thus become one of the research hotspots today. OBJECTIVE： To observe the growth characteristics of human adipose derived stem cells and to test their stem cell features and immunology characters. METHODS： The subcutaneous fat tissue were extracted from three healthy volunteers through liposuction, and then digested with collagenase type I followed by in vitro primary culture and passage culture. Then, the biologica characteristics of passaged cells were observed. Surface antigens and human leukocyte antigens were detected by flow cytometry. And then, peripheral blood lymphocytes were cultured with allogeneic adipose-derived stem cells to observe whether adipose-derived stem cells can accelerate the proliferation of tymphocytes. RESULTS AND CONCLUSION： The harvested adipose-derived stem cells had a strong expansion in vitro with ＂S＂ shape growth curve. The third generation cells were positive for CD29, CD44, CD90 and human leukocyte antigen I, and negative for CD34, CD45, CD31 and human leukocyte antigen I1. Besides, the lymphocytes had no response induced by allogeneic adipose-derived stem cells. The differentiation potential of adipose-derived stem cells used in experiments was confirmed, and they had a weak rejection to the host, which indicates that the adipose-derived stem cells may be a candidate for cell treatment and tissue engineering.