Comparison of cultured mesenchymal stem cells derived from bone marrow or peripheral blood of rats

A. Kamal, D. Iskandriati, I. H. Dilogo, N. Siregar, E. Hutagalung, A. Yusuf, S. Mariya, Kurniadi Husodo
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引用次数: 5

Abstract

Objectives: Mesenchymal stem cells (MSC) are primarily isolated from bone marrow. Peripheral blood is also reported as an alternative source of MSC. This study compared MSC which were isolated and cultured from bone marrow and those from peripheral blood of rats. Methods: MSC from bone marrow and peripheral blood were harvested from 5 male Sprague Dawley rats. After isolation, the cells were grown on tissue culture plates with concentration of 107 cells per well. Observations were conducted to evaluate the attachment of nucleated cells with fibroblast-like morphology. Characterization of MSC was done using reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry assay. Results: The mean number of nucleated cells isolated from the bone marrow on day 0 was higher than those isolated from the peripheral blood. Bone marrow MSC with typical fibroblast-like morphology proliferated rapidly and reached 80% confluency on day 14. Subcultures were able to be conducted on day 15 (first passage) and day 28 (second passage). On days 15 and 28, no nucleated cells remained in peripheral blood cultures. Conclusions: Bone marrow derived MSC are more enriched and grow more efficiently than those derived from peripheral blood.
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大鼠骨髓与外周血间充质干细胞的培养比较
目的:间充质干细胞(MSC)主要从骨髓中分离。外周血也被认为是骨髓间充质干细胞的另一种来源。本研究比较了从大鼠骨髓中分离培养的间充质干细胞和从大鼠外周血中分离培养的间充质干细胞。方法:取5只雄性sd大鼠骨髓和外周血的MSC。细胞分离后,以每孔107个细胞的浓度在组织培养板上生长。观察成纤维细胞样形态有核细胞的附着情况。利用逆转录聚合酶链反应(RT-PCR)和免疫细胞化学方法对MSC进行表征。结果:第0天从骨髓中分离的有核细胞的平均数量高于外周血中分离的有核细胞的平均数量。具有典型成纤维细胞样形态的骨髓间充质干细胞迅速增殖,在第14天达到80%的融合度。传代培养可以在第15天(第一次传代)和第28天(第二次传代)进行。第15天和第28天,外周血培养中没有有核细胞。结论:骨髓来源的间充质干细胞比外周血来源的间充质干细胞更丰富,生长效率更高。
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