本文選題：脂肪干細(xì)胞 + 人纖維細(xì)胞生長(zhǎng)因子10��； 參考：《第二軍醫(yī)大學(xué)》2007年碩士論文

【摘要】： 目的:研究人成纖維細(xì)胞生長(zhǎng)因子10(FGF10)誘導(dǎo)脂肪干細(xì)胞分化的機(jī)制,并為臨床以增脂為主要手段的組織工程和臨床整形美容及創(chuàng)傷修復(fù)提供思路和方法。 方法:1、分離,培養(yǎng)從外科手術(shù)術(shù)中物理分離得到的脂肪干細(xì)胞,并通過(guò)定向分化誘導(dǎo)等方法鑒定其干細(xì)胞特性。2、從人肺組織中抽提總RNA,并利用RT-PCR擴(kuò)增FGF10,構(gòu)建pcDNA3.0-FGF10重組質(zhì)粒,并將其轉(zhuǎn)染至MSC中,利用IHC, ELISA, RT-PCR和western-blotting的方法鑒定pcDNA3.0-FGF10-MSC表達(dá)目的蛋白FGF10的情況。3、利用pcDNA3.0-FGF10-MSC與ADSC體外共培養(yǎng)及裸鼠體內(nèi)注射細(xì)胞的方法證明FGF10的促成脂作用,通過(guò)顯微鏡觀察細(xì)胞改變及RT-PCR和real-time PCR監(jiān)測(cè)ADSC分化過(guò)程中轉(zhuǎn)錄因子的變化。通過(guò)取裸鼠皮下組織切片染色及免疫組化證實(shí)FGF10的體內(nèi)作用。 結(jié)果:1、我們培養(yǎng)的ADSC具有干細(xì)胞特性,并能多向誘導(dǎo)分化。2、構(gòu)建的pcDNA3.0-FGF10-MSC表達(dá)載體能持續(xù)表達(dá)目的蛋白FGF10。3、體外共培養(yǎng)實(shí)驗(yàn)發(fā)現(xiàn)ADSC細(xì)胞內(nèi)脂滴增多,分化轉(zhuǎn)錄因子表達(dá)增高。裸鼠體內(nèi)實(shí)驗(yàn)發(fā)現(xiàn)皮下脂肪層明顯增厚。 結(jié)論:FGF10能在體外及體內(nèi)促進(jìn)脂肪干細(xì)胞的成熟分化。
[Abstract]:Objective: to study the mechanism of adipose stem cell differentiation induced by human fibroblast growth factor (FGF10), and to provide ideas and methods for clinical tissue engineering, cosmetic surgery and wound repair.Methods the adipose stem cells were isolated and cultured during surgery. The stem cell characteristics of the stem cells were identified by directional differentiation induction. The total RNAs were extracted from human lung tissue. FGF10 was amplified by RT-PCR, and the recombinant plasmid of pcDNA3.0-FGF10 was constructed.The expression of pcDNA3.0-FGF10-MSC protein FGF10 was identified by IHC, Elsa, RT-PCR and western-blotting. The effect of FGF10 on lipid production was proved by co-culture of pcDNA3.0-FGF10-MSC and ADSC in vitro and injection of cells in nude mice.The changes of transcription factors in the differentiation of ADSC were observed by microscope and the changes of transcription factors during ADSC differentiation were monitored by RT-PCR and real-time PCR.The effect of FGF10 in vivo was confirmed by immunohistochemical staining and staining of subcutaneous tissue sections of nude mice.Results: the ADSC cultured in vitro had stem cell characteristics and was able to induce differentiation. The constructed pcDNA3.0-FGF10-MSC expression vector could continuously express the target protein FGF10.3. In vitro co-culture experiments showed that the lipid droplets in ADSC cells increased and the expression of differentiation transcription factors increased.The subcutaneous adipose layer was obviously thickened in nude mice.Conclusion: FGF10 can promote the maturation and differentiation of adipose stem cells in vitro and in vivo.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：R329

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