中国组织工程研究与临床康复2011，Vol.15Issue(10)：1750-1754,5.DOI:10.3969/j.issn.1673-8225.2011.10.009
长期培养人脐带间充质干细胞的生物活性及其限制性
Biological activity and restriction of human umbilical cord mesenchymal stem cells during long-term culture in vitro
许超 1廖继东 1柳菁 2毛文哲 1李扬秋3
作者信息
- 1. 暨南大学医学院田家炳医学实验中心,广东省广州市,510632
- 2. 暨南大学医学院生化教研室,广东省广州市,510632
- 3. 暨南大学医学院血液病研究所,广东省广州市,510632
- 折叠
摘要
Abstract
BACKGROUND:Mesenchymal stem cells (MSCs) possess multi-lineage differentiation potential and immunomodulatory effects,and can be made long-term culture in vitro.However,it is still not very clear about the restriction and biological activity feature of MSCs during long-term cultu re in vitro.OBJ ECTIVE: To investigate changes of biological activity feature and restriction of human umbilical cord MSCs (hUC-MSCs) of long-term culture in vitro.METHODS: The umbilical cord from cesarean section at term newborns was collected in sterile condition. MSCs were isolated from human umbilical cord by collagenase Ⅱ digestion. The hUC-MSCs were obtained by plastic surface adhesion selection,continuous passage culture by trypsin digestion. The cells surface antigens were identified by flow cytometry, adipogenic and osteogenic ability were determined by specially inducing culture from third generation. A series of different generation cells were collected for following analysis. The cell growth curve was analyzed by MTT assay. The cell cycle and apoptosis rate were measured by flow cytometry. The cell adhesive ability was detected by piece of climbing.RESULTS AND CONCLUSION: The cultured hUC-MSCs phenotype was CD105+/CD29+/CD44+/CD31-/CD34-/CD45-/HLADR-,and could were induced toward osteoblast and adipocyte differentiation under appropriate experimental conditions. There was no significant change in the cellular morphology and the cell growth curve of subculture 3-23 generation. After subculture 28 generation, cell growth became slowly. When subcultu re 33 generation was compared with subculture 3 generation, the cellular number of G0/G1 phase was decrease 17.9％ , and S+G2/M phase and the cell apoptotic rate were increase 103.4％ and 316.7％ respectively. Their diversity was statistical significant (P ＜ 0.01). Within subculture 3-18 generation, their diversity of the cellular number of G0/G1 and S+G2/M phase and the cell apoptotic rate were not statistical significant (P ＞ 0.05). Our data show that following increase of subculture frequency, hUC-MSCs proliferate and adherent capabilities of were decrease, while its apoptosis rate increased, in condition of long-term culture in vitro. lt's all biological activity has gradually decreasing trend. The best biological activity period appeared in 3 to 18 generations.关键词
人脐带间充质干细胞/培养/生物活性/限制性/脐带分类
医药卫生引用本文复制引用
许超,廖继东,柳菁,毛文哲,李扬秋..长期培养人脐带间充质干细胞的生物活性及其限制性[J].中国组织工程研究与临床康复,2011,15(10):1750-1754,5.
