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利用骨髓间充质干细胞自分泌的细胞外基质培养间充质干细胞的生物安全性

The biological safety evaluation for bone marrow mesenchymal stem cells cultured on decellularized extracellular matrix
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摘要 目的 评价利用骨髓间充质干细胞(BM-MSCs)自分泌的细胞外基质(ECM)培养MSCs的生物安全性。方法 通过对ECM培养的BM-MSCs细胞上清液行病原学、内毒素及血清残留量检测,同时采用G显带方法对细胞染色体进行核型分析和软琼脂培养克隆形成试验。结果 利用ECM培养第5代BM-MSCs的上清液各种病原学检测均为阴性,内毒素(〈0.12 EU/mL)及血清残留量(〈2.5 ng/mL)检测符合生物学制剂的限制标准。第10代的细胞染色体核型稳定,无异常改变和致瘤性。结论 利用ECM培养的BM-MSCs具有良好的生物安全性,为下一步临床应用提供了依据。 Objective To investigate the biological safety of bone marrow mesenchymal stem cells (BM-MSCs) cultured on decellularized extracellular matrix (ECM). Methods Culture medium of BM-MSCs cultured was collected for detection of bacteria, virus, endotoxin and residual serum. The Gelclot semiquantitative method was adopted to analyze the content of bacterial endotoxin according to China Pharmacopoeia. Residual serum was measured by ELISA kit. The oncogenicity of BM- MSCs was verified by soft agar cloning method and the genetic stability of BM-MSCs was examined by karyotype analysis. Results Bacteria and virus were not detected in cell supernatant of BM- MSCs cultured on ECM; content of bacterial endotoxin (〈0.12 EU/mL) and residual serum (〈2.5 ng/mL) met certain criteria according to China Phaimacopoeia. The tenth passage BM-MSCs maintained normal karyotype and had no tumorigenicity by soft agar cloning test. Conclusions BM-MSCs cultured on ECM had good biological safety. The present study indicates that decellularized BM- MSCs had considerable potential clinical application.
作者 何宏亮 詹玛琍 陈兆武 徐少保 李宜 HE Hong-liang1, ZHAN Ma-li2, CHEN Zhao-wu3, XU Shao-bao1, LI Yi1 (1. Department of lnfectious Diseases, Anhui Provincial Hospital, Anhui Medical University, Hefei, Anhui 230001 ; 2. Department of Pathology, Anhui Provincial Hospital, Anhui Medical University, Hefei ,Anhui 230001 ; 3. Department of Laboratory ,Anhui Provincial Cancer Hospital, Hefei, Anhui 230031, China )
出处 《热带医学杂志》 CAS 2017年第12期1577-1579,1588共4页 Journal Of Tropical Medicine
关键词 间充质干细胞 生物安全性 染色体核型 Mesenchymal stem cells Biological safety evaluation Chromosome karyotype
作者简介 何宏亮(1983-),医学博士,主治医师,研究方向:干细胞治疗肝衰竭的基础与临床,E—mail:44271370@qq.com
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