改良全骨髓贴壁法培养SD大鼠骨髓间充质干细胞及其生物学特性

doi:10.3969/j.issn.1006-5725.2025.24.009

Abstract

Abstract:

Objective To establish an efficient method for the isolation， culture， and purification of rat bone marrow mesenchymal stem cells （BMSCs）， while simultaneously investigating the biological characteristics of these cells. Methods The 3 ~ 4 weeks old male SD rats were sacrificed via cervical dislocation， followed by the sterile dissection of both femurs and tibiae. BMSCs from rats were separated and cultivated employing two techniques： the traditional marrow flush-out method and the optimized whole marrow adhesion method. When cell confluence reached 80%， the cells were digested for subculture and purified. Cellular morphology was observed under an inverted microscope； Colony formation and the colony-forming rate of BMSCs were assessed following Giemsa staining； Flow cytometric analysis was performed to identify specific surface markers （CD29 and CD45）； Cell proliferation capacity was measured using the CCK-8 assay， and growth curves from day 1 to day 8 after passaging were plotted； To assess osteogenic differentiation capability， BMSCs were directed toward the osteoblast lineage， and the differentiation efficacy was analyzed in the two groups through ALP staining with quantitative analysis and Alizarin Red staining followed by quantification； Additionally， adipogenic differentiation was induced in BMSCs， and the lipid accumulation capacity of the two groups was detected via Oil Red O staining and subsequent quantitative measurement. Results Compared to the control group， SD rat BMSCs isolated and purified in vitro using the modified and optimized whole bone marrow adherence method exhibited superior morphology， higher purity， and enhanced proliferation capacity. Furthermore， regarding cellular biological properties， BMSCs from the modified group showed a slight enhancement in osteogenic differentiation potential and a mild reduction in adipogenic differentiation capacity compared to those from the control group. Conclusion BMSCs derived from SD rats using the optimized whole marrow adhesion technique demonstrate superior morphological characteristics， enhanced proliferation ability， and optimal differentiation capability， thus qualifying as viable progenitor cells for tissue engineering purposes.

Key words: bone marrow mesenchymal stem cells, modified and optimized whole marrow adhesion method, biological characteristics, differentiation

CLC Number:

R-331

Yusheng HUANG,Xinyu LIU,Suishan. ZHOU. Study on the culture and biological characteristics of SD rat bone marrow mesenchymal stem cells using optimized whole marrow adhesion method[J]. The Journal of Practical Medicine, 2025, 41(24): 3860-3866.

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References 26

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Study on the culture and biological characteristics of SD rat bone marrow mesenchymal stem cells using optimized whole marrow adhesion method

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