【摘要】：目的 觀察成骨生長(zhǎng)肽(OGP)在成脂及成骨性培養(yǎng)基中對(duì)大鼠骨髓間充質(zhì)干細(xì)胞(MSC)增殖分化的影響。 方法 Percoll法分離大鼠骨髓間充質(zhì)干細(xì)胞,通過頻繁更換培養(yǎng)液及反復(fù)傳代的方法純化細(xì)胞。用傳至2代細(xì)胞作MTT實(shí)驗(yàn),以此確定OGP對(duì)MSC起作用的最大活性濃度;傳3代MSC接種于6孔培養(yǎng)板上,共接種13板,分為成脂、成骨誘導(dǎo)兩組,每板細(xì)胞都分為實(shí)驗(yàn)組和對(duì)照組,對(duì)照組不含成骨生長(zhǎng)肽,實(shí)驗(yàn)組含有10~(-11)mol/L的成骨生長(zhǎng)肽。脂肪細(xì)胞鑒定:成脂誘導(dǎo)7、14、21天各取1板細(xì)胞做實(shí)驗(yàn),用TG試劑盒在全自動(dòng)生化分析儀上測(cè)定細(xì)胞中甘油三酯的總量;成脂誘導(dǎo)21天后取預(yù)置蓋玻片的1板細(xì)胞行脂肪細(xì)胞油紅O染色。成骨細(xì)胞鑒定:取成骨誘導(dǎo)8、12、16天細(xì)胞各2板,分別用ALP試劑盒、鈣試劑盒測(cè)定細(xì)胞內(nèi)堿性磷酸酶活性及每孔鈣含量;成骨誘導(dǎo)30天后取預(yù)置蓋玻片的1板細(xì)胞,做Von Kossa染色,計(jì)算每張玻片鈣結(jié)節(jié)數(shù)。 結(jié)果 傳3代大鼠骨髓間充質(zhì)干細(xì)胞成脂誘導(dǎo)7天、14天、21天后分化為脂肪細(xì)胞的陽(yáng)性率以及甘油三酯含量,實(shí)驗(yàn)組均高于對(duì)照組;成骨誘導(dǎo)8天、12天、16天后細(xì)胞內(nèi)堿性磷酸酶活性測(cè)定以及鈣含量測(cè)定,實(shí)驗(yàn)組均高于對(duì)照組,30天后礦化結(jié)節(jié)的形成,實(shí)驗(yàn)組高于對(duì)照組。 結(jié)論 隨著培養(yǎng)條件不同,成骨生長(zhǎng)肽可促進(jìn)大鼠骨髓間充質(zhì)干細(xì)胞向脂肪細(xì)胞或成骨細(xì)胞分化。
[Abstract]:Objective to observe the effect of osteogenic growth peptide (OGP) on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (MSC) in adipogenic and osteoblast medium. Methods Rat bone marrow mesenchymal stem cells were isolated by Percoll method. The maximum active concentration of OGP on MSC was determined by the MTT assay of passage 2 cells, 13 plates were inoculated on 6 well culture plate, 13 plates were inoculated on 6 well culture plate, each plate cells were divided into experimental group and control group, and each of them was divided into experimental group and control group. The control group did not contain osteogenic growth peptide, while the experimental group contained 10 ~ (- 11) mol/L osteogenic growth peptide. Adipocyte identification: 1 plate cell was taken for 21 days after adipogenic induction, and the total triglyceride content was measured by TG kit in automatic biochemical analyzer. After 21 days of adipogenic induction, 1 plate cells prepositioned with cover glass were stained with fat cell oil red O. Osteoblast identification: two plates of osteoblasts were taken from each of the cells on day 8, 12 and 16, respectively. The activity of alkaline phosphatase and the content of calcium in each hole were measured by ALP kit and calcium kit respectively. After 30 days of osteogenesis induction, the 1 plate cells prepositioned with glass slide were stained with Von Kossa. Calculate the number of calcium nodules per slide. Results the positive rate and triglyceride content of adipocytes differentiated into adipocytes from rat bone marrow mesenchymal stem cells induced by adipogenic induction for 7 days and 14 days and 21 days later were higher in the experimental group than in the control group. The activity of alkaline phosphatase and the content of calcium in the cells of the experimental group were higher than those of the control group after 30 days of osteogenic induction, and the formation of mineralized nodules in the experimental group was higher than that in the control group. Conclusion osteoblast growth peptide can promote the differentiation of rat bone marrow mesenchymal stem cells into adipocytes or osteoblasts with different culture conditions.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R329.2

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