本文選題：成骨細(xì)胞　切入點(diǎn)：骨髓間充質(zhì)干細(xì)胞　出處：《遼寧醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的 通過應(yīng)用Transwell培養(yǎng)板，將SD乳鼠顱蓋骨成骨細(xì)胞與乳兔骨髓間充質(zhì)干細(xì)胞（BMSCs）共培養(yǎng)，觀察共培養(yǎng)環(huán)境下SD乳鼠顱蓋骨成骨細(xì)胞對乳兔BMSCs向成骨細(xì)胞方向的誘導(dǎo)作用。 方法 取出生3天SD大鼠乳鼠顱蓋骨，采用組織塊培養(yǎng)法培養(yǎng)成骨細(xì)胞；取乳兔長骨，采用全骨髓培養(yǎng)法培養(yǎng)BMSCs，并用流式細(xì)胞儀分析細(xì)胞表面CD44、CD34、CD45、CD90的表達(dá)對其進(jìn)行鑒定；應(yīng)用Transwell雙層細(xì)胞培養(yǎng)板將大鼠成骨細(xì)胞與兔骨髓間充質(zhì)細(xì)胞共培養(yǎng)，實(shí)驗(yàn)組于培養(yǎng)上室接種成骨細(xì)胞，對照組培養(yǎng)上室不接種細(xì)胞，兩組的培養(yǎng)下室均接種BMSCs。觀察各組BMSCs形態(tài)的變化，，用酶標(biāo)儀檢測誘導(dǎo)分化后BMSCs的ALP活性，RT-PCR法分別檢測兩組BMSCs的骨鈣素、骨形態(tài)發(fā)生蛋白-2（BMP-2）、Ⅰ型膠原基因的表達(dá)情況。 結(jié)果 1、大鼠成骨細(xì)胞與兔BMSCs共培養(yǎng)后，實(shí)驗(yàn)組BMSCs形態(tài)逐漸向成骨樣細(xì)胞轉(zhuǎn)化，而對照組無明顯變化； 2、實(shí)驗(yàn)組BMSCs的ALP表達(dá)活性為0.365±0.016，對照組為0.17±0.012，即實(shí)驗(yàn)組BMSCs中ALP活性高于對照組。 3、實(shí)驗(yàn)組誘導(dǎo)后的BMSCs的骨鈣素、BMP-2、Ⅰ型膠原基因有所表達(dá)，對照組無表達(dá)。 結(jié)論 乳鼠成骨細(xì)胞與乳兔BMSCs共培養(yǎng)后，能夠誘導(dǎo)BMSCs向成骨細(xì)胞分化，BMSCs表達(dá)了成骨細(xì)胞的特異性表面標(biāo)志。
[Abstract]:PurposeThe osteoblasts from calvaria of neonatal SD rats and bone marrow mesenchymal stem cells (BMSCs) of neonatal SD rats were co-cultured with Transwell culture plate to observe the induction of osteoblasts from calvaria to osteoblasts of newborn rabbits in co-cultured environment.MethodThe cranial calvaria of SD rats was taken out for 3 days and osteoblasts were cultured by tissue mass culture, and BMSCs were cultured by whole bone marrow culture method.Rat osteoblasts and rabbit bone marrow mesenchymal cells were co-cultured with Transwell double cell culture plate. The experimental group was inoculated with osteoblasts in the upper chamber, but the control group was not inoculated in the upper chamber.The morphological changes of BMSCs in each group were observed. The expression of osteocalcin, bone morphogenetic protein (BMP)-2 BMP-2 and type I collagen gene in BMSCs were detected by reverse transcription-polymerase chain reaction (RT-PCR) for detecting the ALP activity of BMSCs induced by differentiation.Result1. After co-culture of rat osteoblasts with rabbit BMSCs, the morphology of BMSCs in the experimental group was gradually transformed to osteoblast-like cells, while in the control group, there was no significant change.2. The ALP expression activity of BMSCs was 0.365 鹵0.016 in the experimental group and 0.17 鹵0.012 in the control group. That is, the ALP activity in the BMSCs of the experimental group was higher than that in the control group.3. Osteocalcin 2, type I collagen gene in BMSCs was expressed in the experimental group, but not in the control group.ConclusionAfter co-culture of neonatal rat osteoblasts and BMSCs, BMSCs could be induced to differentiate into osteoblasts and express the specific surface markers of osteoblasts.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329

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