本文選題：人胚胎生殖腺嵴和腸背系膜來源成纖維樣細胞 + 人胚胎生殖細胞��； 參考：《吉林大學(xué)》2007年博士論文

【摘要】： 為優(yōu)化人胚胎生殖細胞(hEG)的體外培養(yǎng)體系及研究5-氮雜胞苷對hEG向心肌前體細胞分化的影響,我們分離培養(yǎng)了人胚胎生殖腺嵴和腸背系膜中原始生殖細胞(PGCs)周圍的成纖維樣細胞,證實其具有穩(wěn)定的生物學(xué)特性,并首次證實其表達體外培養(yǎng)hEG所必需的細胞因子。以此人胚胎成纖維樣細胞作為飼養(yǎng)層細胞,模擬PGCs在早期胚胎發(fā)育過程中不斷遷移、擴增并保持未分化狀態(tài)時的體內(nèi)環(huán)境,同時添加bFGF、LIF和forskolin細胞因子,體外成功培養(yǎng)hEG,并對其生物學(xué)特性進行鑒定。所培養(yǎng)的hEG可傳8～10代,并能夠自我更新,保持未分化狀態(tài)、正常人類二倍體核型及多向分化能力,建立了一種新的hEG體外培養(yǎng)方法。為hEG的進一步研究及其臨床應(yīng)用奠定了基礎(chǔ)。在此基礎(chǔ)上,進一步利用半定量逆轉(zhuǎn)錄多聚酶鏈反應(yīng)檢測5-氮雜胞苷對hEG向心肌前體細胞分化的影響。初步研究表明,5-氮雜胞苷可能具有促進hEG向心肌前體細胞分化的能力。同時,我們亦自人早期胚胎中分離培養(yǎng)人胚胎心肌細胞,并對其生物學(xué)特性進行初步研究。為今后心血管疾病的干細胞移植治療研究奠定了基礎(chǔ)。
[Abstract]:In order to optimize the culture system of human embryonic germ cells (EGG) in vitro and to study the effect of 5-azacytidine on the differentiation of hEG into cardiomyocyte precursor cells,We isolated and cultured fibroblasts around the primordial germ cells (PGCs) of human embryonic gonadal ridge and dorsal mesenteric membrane, and confirmed their stable biological characteristics, and demonstrated for the first time the expression of cytokines necessary for the culture of hEG in vitro.Human embryonic fibroblast cells were used as feeder layer cells to mimic the migration of PGCs during early embryonic development, to amplify and maintain the undifferentiated environment in vivo, and to add bFGF LIF and forskolin cytokines at the same time.HEGG was successfully cultured in vitro and its biological characteristics were identified.The cultured hEG can be transferred for 810 generations, and can renew itself, maintain undifferentiated state, normal human diploid karyotype and multidirectional differentiation ability. A new method of hEG culture in vitro was established.It lays a foundation for the further study and clinical application of hEG.On this basis, semi-quantitative reverse transcription polymerase chain reaction was used to detect the effect of 5-azacytidine on the differentiation of hEG into cardiomyocyte precursor cells.The preliminary study showed that 5-azacytidine might promote the differentiation of hEG into cardiomyocyte precursor cells.At the same time, human embryonic cardiomyocytes were isolated from human early embryos and their biological characteristics were studied.It will lay a foundation for the research of stem cell transplantation therapy for cardiovascular disease in the future.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2007
【分類號】：R329

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本文編號：1749133