發(fā)布時(shí)間：2018-01-28 04:31

  本文關(guān)鍵詞： 電磁場 骨髓間充質(zhì)干細(xì)胞 成骨　出處：《浙江大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：實(shí)驗(yàn)?zāi)康模簽榱搜芯康皖l電磁場(EMF)對(duì)大鼠骨髓間充質(zhì)干細(xì)胞(BMSCs)分化、增殖、細(xì)胞周期和體內(nèi)作用的效果。 實(shí)驗(yàn)方法：將原代的骨髓間充質(zhì)干細(xì)胞培養(yǎng)至第三代,分成四個(gè)組別。輻照組細(xì)胞被置于0.5mT,50HZ的電磁場中；實(shí)驗(yàn)組細(xì)胞被置于含成骨誘導(dǎo)液的培養(yǎng)基中；聯(lián)合組細(xì)胞被置于含成骨誘導(dǎo)液的培養(yǎng)基同時(shí)接受0.5mT,50HZ的電磁場的輻照。MTT法被用于估計(jì)細(xì)胞的增殖情況,堿性磷酸酶活性檢測用于估計(jì)細(xì)胞的分化情況。分別在實(shí)驗(yàn)的第3天、第7天和第10天檢測這兩個(gè)指標(biāo)。在第7天檢測細(xì)胞周期的改變情況,在第12天檢測骨結(jié)節(jié)的形成情況。此外,在第10天用RT-PCR檢測Ⅰ型膠原的基因表達(dá)。輻照組和對(duì)照組的細(xì)胞分別被植入相同的PLGA支架后再分別植入到不同的大鼠股骨皮質(zhì)中。分別在第4、第8周分別取材行病理切片并HE染色后觀察成骨結(jié)果。 實(shí)驗(yàn)結(jié)果：結(jié)果顯示低頻電磁場干預(yù)后加快了干細(xì)胞的增殖(p0.05),提高了干細(xì)胞的分化(p0.05),使G2/M+S期的細(xì)胞比例顯著升高。與空白組相比,EMF輻照組有更顯著的Ⅰ型膠原相關(guān)mRNA得以表達(dá)(p0.05)。EMF聯(lián)合成骨誘導(dǎo)液組形成了明顯的骨結(jié)節(jié)。在輻照組體內(nèi)實(shí)驗(yàn)中,HE染色后觀察到了明顯的骨組織形成。 結(jié)論：低頻電磁場(0.5mT,50HZ)能顯著促進(jìn)骨髓間充質(zhì)干細(xì)胞的增殖和分化。
[Abstract]:Aim: to study the effects of low frequency electromagnetic field (EMF) on differentiation, proliferation, cell cycle and in vivo action of rat bone marrow mesenchymal stem cells (BMSCs). Methods: primary bone marrow mesenchymal stem cells were cultured to the third generation and divided into four groups. The cells of the experimental group were placed in the medium containing osteogenic medium. The combined group of cells were placed in the medium containing osteoblast and exposed to the electromagnetic field of 0.5mTX 50HZ. MTT method was used to estimate the proliferation of the cells. Alkaline phosphatase activity was used to estimate cell differentiation. These two indexes were detected on day 3, day 7 and day 10, respectively. Changes in cell cycle were detected on day 7. The formation of bone nodules was detected on the 12th day. On the 10th day, the gene expression of type I collagen was detected by RT-PCR. The cells of irradiation group and control group were implanted with the same PLGA scaffold, and then implanted into different femur cortex of rats. 4. At the 8th week, the osteogenesis was observed by pathological sections and HE staining. The results showed that the low frequency electromagnetic field intervention accelerated the proliferation of stem cells and increased the differentiation of stem cells. The proportion of cells in G2 / MS phase was significantly increased compared with the blank group. The expression of type I collagen-associated mRNA in the EMF irradiation group was more significant, and the bone nodules were formed in the combined osteogenic inducer group. In vivo experiments were carried out in the irradiated group. The formation of bone tissue was observed after HE staining. Conclusion: low frequency electromagnetic field can significantly promote the proliferation and differentiation of bone marrow mesenchymal stem cells.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R329

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