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短期凍存兔骨髓間充質(zhì)干細(xì)胞對(duì)其生物特性的影響

發(fā)布時(shí)間:2018-12-09 08:57
【摘要】:目的探討短期凍存對(duì)兔骨髓間充質(zhì)干細(xì)胞(BMSCs)的增殖、生物特征及成骨能力的影響。方法選取第5代兔BMSCs凍存,30d后復(fù)蘇、體外培養(yǎng)至第10代作為實(shí)驗(yàn)組,設(shè)相同代數(shù)未凍存的BMSCs為對(duì)照組。MTT、細(xì)胞周期、細(xì)胞表面標(biāo)志及成骨誘導(dǎo)分化后堿性磷酸酶(ALP)水平、Ⅰ型膠原及鈣結(jié)節(jié)染色作為其增殖、生物特征及成骨能力的評(píng)價(jià)指標(biāo)。結(jié)果凍存后BMSCs生長(zhǎng)潛伏期延長(zhǎng),但生長(zhǎng)狀態(tài)無(wú)明顯影響;增殖指數(shù)和G1期細(xì)胞所占比例分別為42.9%±3.4%、57.0%±3.4%,細(xì)胞表面標(biāo)志CD44陽(yáng)性率為93.62%±1.05%,不表達(dá)CD45;成骨誘導(dǎo)第7、14天ALP水平(U/gprot)分別為6.73±1.92、15.99±4.36,第14、21天,Ⅰ型膠原、鈣結(jié)節(jié)茜素紅染色陽(yáng)性;以上結(jié)果與對(duì)照組比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論短期凍存對(duì)兔BMSCs增殖、生物特征及成骨分化潛能無(wú)明顯影響,凍存后可繼續(xù)進(jìn)一步研究。
[Abstract]:Objective to investigate the effects of short-term cryopreservation on (BMSCs) proliferation, biological characteristics and osteogenesis of rabbit bone marrow mesenchymal stem cells (BMSCs). Methods the fifth generation rabbit BMSCs was cryopreserved for 30 days, then resuscitated and cultured in vitro to the 10th passage as the experimental group. The BMSCs with the same algebra was used as the control group. The MTT, cell cycle was used as the control group. Cell surface markers and alkaline phosphatase (ALP) levels after osteogenic differentiation, type I collagen and calcium nodule staining were used as indicators of proliferation, biological characteristics and osteogenesis ability. Results the incubation period of BMSCs was prolonged after cryopreservation, but the growth state was not affected. The percentage of proliferative index and G1 phase cells were 42.9% 鹵3.4% 鹵3.4% and 57.0% 鹵3.4%, respectively. The positive rate of CD44 on cell surface marker was 93.62% 鹵1.05%, and CD45; was not expressed. The level of ALP (U/gprot) was 6.73 鹵1.92 鹵4.36 on the 14th day after osteogenesis induction and 15.99 鹵4.36 on the 14th day after osteogenesis induction, and the type 鈪,

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