目的 研究人脂肪来源间充质干细胞(human adipose-derived stem cells, hADSCs)的制备及其质量检验方法 , 为临床研究提供安全合格的干细胞。方法 取50例人吸脂术抽取的脂肪, 剪碎, 用胰酶和胶原酶消化、分离、培养, 并按照《干细胞制剂质量控制及临床前研究指导原则》对其进行细胞形态、数量、活率、无菌、支原体、人源特定病毒及猪源病毒、内毒素、免疫抑制活性、分化能力、免疫表型等检测及染色体核型。结果 50例hADSCs;细菌、内毒素检测阴性, 无内外源致病菌;干细胞免疫表型检测CD90、CD105表达阳性, 阳性率＞95%, CD34 、CD45和HLA-DR表达阴性, 阳性率<2%;具有成骨、成脂分化潜能;能抑制异体淋巴细胞的增殖。间充质干细胞活性冻存前≥92％, 冻存复苏后≥82％。结论 按照本工艺及标准制备的hADSCs符合质量控制标准, 为同类干细胞制备及其检定过程标准化提供了试验依据。

Objective To study the human adipose derived mesenchymal stem cells in vitro amplification and quality inspection Methods and provide safe and qualified stem cells for clinical research. Methods Fifty human source liposuctioned fat, sheared, underwent trypsin and collagenase digestion, separation, culture, and according to the “stem cell-based medicinal product quality control of pre clinical research guiding principle” to detect the cell morphology, quantity, live rate, sterilization, mycoplasma, human source specific virus and swine virus, endotoxin, immunosuppressive activity, differentiation ability, immunophenotype detection and chromosome karyotype. Results Human adipose derived mesenchymal stem cell activity before cryopreservation is ≥92%, after revival≥82%; bacteria and endotoxin test negative, no internal and external source pathogenic bacteria; stem cell immunophenotype detection of CD90, CD105 positive, the positive rate>95%, CD34, CD45 and HLA-DR negative expression, the positive rate<2%; with osteogenic and adipogenic differentiation proficiency , it can inhibit the proliferation of allogeneic lymphocytes. Conclusions The preparation process and the standard of human adipose derived mesenchymal stem cell line according to the quality control standard of “stem cell-based medicinal product quality control of pre clinical research guiding principle”, it provides the experimental basis for the similar stem cell preparation and standardizing verification process.