本文選題：臍血 + 間充質(zhì)干細(xì)胞��； 參考：《四川大學(xué)》2006年碩士論文

【摘要】：目的： 研究凍存復(fù)蘇對人臍血間充質(zhì)干細(xì)胞(UCBMSCs)生長狀況的影響；研究復(fù)蘇后的UCBMSCs在肝細(xì)胞生長因子(HGF)和成纖維細(xì)胞生長因子-4(FGF-4)的聯(lián)合誘導(dǎo)下，向肝細(xì)胞分化的潛能，以及類肝細(xì)胞對UCBMSCs是否具有定向誘導(dǎo)的作用。 方法： 1．用含10％二甲基亞砜(DMSO)的細(xì)胞培養(yǎng)基將第6代UCBMSCs凍存于液氮中；6個月后復(fù)蘇細(xì)胞，觀察細(xì)胞的生長形態(tài)，臺盼藍(lán)染色排除法計(jì)數(shù)活細(xì)胞。 2．在復(fù)蘇后培養(yǎng)至第8代的細(xì)胞中，加入40ng/ml HGF和10ng／mlFGF-4，陰性對照組為不加誘導(dǎo)因子培養(yǎng)的同代細(xì)胞，，陽性對照組為HepG2；于加入誘導(dǎo)因子后第7、14、21及28天檢測細(xì)胞胞漿內(nèi)CK818、白蛋白及糖原的表達(dá)。 3．將類肝細(xì)胞與新鮮制備的UCBMSCs共培養(yǎng)；將與類肝細(xì)胞同期培養(yǎng)的未誘導(dǎo)細(xì)胞和新鮮制備的UCBMSCs共培養(yǎng)作為陰性對照組。于28天后檢測UCBMSCs中，CK818和白蛋白的雙表達(dá)以及糖原的表達(dá)。
[Abstract]:Objective:To study the effect of cryopreservation and resuscitation on the growth of human umbilical cord blood mesenchymal stem cells (UCB MSCs), and to study the potential of UCBMSCs after resuscitation to differentiate into hepatocytes induced by the combination of hepatocyte growth factor (HGF) and fibroblast growth factor (FGF-4).And whether hepatoid cells have directional induction to UCBMSCs.Methods:1.The sixth passage UCBMSCs was frozen in liquid nitrogen on the cell culture medium containing 10% dimethyl sulfoxide (DMSO). After 6 months the cells were resuscitated to observe the growth morphology of the cells and the living cells were counted by trypan blue staining exclusion method.2.After resuscitation, 40ng/ml HGF and 10 ng / ml FGF-4 were added to the cells cultured in the 8th passage. The negative control group was the same generation of cells cultured without inducible factor.The expression of CK818, albumin and glycogen in the cytoplasm of the cells was detected on the 21st and 28th day after the induction factor was added to the positive control group.3.Hepatoid cells were co-cultured with freshly prepared UCBMSCs and uninduced cells and freshly prepared UCBMSCs were co-cultured as negative control group.After 28 days, the double expression of CK818 and albumin and the expression of glycogen in UCBMSCs were detected.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R329.2

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