目的 建立大鼠骨髓间充质干细胞（BMSCs）高效的分离、培养和纯化的方法，同时探究BMSCs的生物学特性。 方法 取3 ~ 4周龄的SD雄性大鼠脱臼处死，在无菌条件下分离双侧股骨、胫骨，分别采用传统冲髓法和改良全骨髓贴壁法分离培养大鼠BMSCs，待细胞融合率达80%时，进行消化传代及纯化培养。倒置显微镜下观察细胞形态；吉姆萨染色后观察BMSCs克隆形成情况及克隆形成率；流式细胞仪检测细胞表面特异抗原（CD29和CD45）；利用CCK-8检测细胞增殖能力，同时绘制细胞传代后第1 ~ 8天的生长曲线；诱导BMSCs向成骨细胞分化，分别利用碱性磷酸酶（ALP）染色、定量及茜素红染色、定量方法，对两组BMSCs的成骨分化能力进行检测；另一方面，诱导BMSCs向脂肪细胞分化，利用油红O染色及定量方法，对两组BMSCs的成脂分化能力进行检测。 结果 相较于对照组，应用改良全骨髓贴壁培养法体外分离、纯化培养的SD大鼠BMSCs形态好、纯度高、增殖能力强。同时，在细胞生物学特性方面，改良组BMSCs较对照组BMSCs的成骨分化能力轻微增强，成脂分化能力轻微减弱。 结论 改良全骨髓贴壁法分离培养的SD大鼠BMSCs，其细胞形态好、增殖能力强、分化能力优，可以选择作为组织工程的种子细胞。

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.