發(fā)布時(shí)間：2019-04-29 07:18

【摘要】：自從1998年Thomson和Gearhart分別從胚泡內(nèi)細(xì)胞群和原始生殖細(xì)胞成功地建立了人類胚胎干細(xì)胞系以來(lái),干細(xì)胞研究已成為當(dāng)今生命科學(xué)領(lǐng)域中的前沿和熱點(diǎn)。研究?jī)?nèi)容主要涉及胚胎干細(xì)胞的培養(yǎng)建系、誘導(dǎo)干細(xì)胞向特定組織細(xì)胞的分化及在生物和醫(yī)學(xué)中的應(yīng)用。來(lái)源于原始生殖細(xì)胞(Primordial Germ Cell,PGC)的胚胎干細(xì)胞(embryonic stem cell,ESC),又稱為胚芽干細(xì)胞(Embryonic Germ Cell,EGC),具有與內(nèi)細(xì)胞群來(lái)源的胚胎干細(xì)胞相同的生物學(xué)特性。本文系統(tǒng)地研究了小鼠胚芽干細(xì)胞的提取、體外擴(kuò)增;胚芽干細(xì)胞向肝細(xì)胞的定向誘導(dǎo)分化;和胚芽干細(xì)胞肝內(nèi)移植的實(shí)驗(yàn)研究。 一、胚芽干細(xì)胞的提取和體外擴(kuò)增: 胚芽干細(xì)胞的提取和培養(yǎng)是進(jìn)行深入研究的前提,我們首先進(jìn)行了培養(yǎng)體系的篩選(見(jiàn)系列論文的第一部分)。 胚芽干細(xì)胞與來(lái)源于內(nèi)細(xì)胞群的胚胎干細(xì)胞一樣,在體外培養(yǎng)過(guò)程中,極易發(fā)生分化。為了在體外維持胚胎干細(xì)胞的增殖和未分化狀態(tài),建立適宜的培養(yǎng)體系是關(guān)鍵。胚胎干細(xì)胞對(duì)體外培養(yǎng)條件要求非常嚴(yán)格,不同種屬動(dòng)物的胚胎干細(xì)胞要求的培養(yǎng)條件也不同。近年來(lái)對(duì)來(lái)源于內(nèi)細(xì)胞群的胚胎干細(xì)胞的培養(yǎng)條件和方法進(jìn)行了廣泛的研究,積累了豐富的經(jīng)驗(yàn)。但是有關(guān)胚芽干細(xì)胞的培養(yǎng)方法報(bào)道較少。本研究對(duì)適用于昆明種小鼠胚芽干細(xì)胞的體外培養(yǎng)體系進(jìn)行了篩選。觀察了基礎(chǔ)培養(yǎng)液、添加白血病抑制因子、同種胎鼠成纖維細(xì)胞條件培養(yǎng)液、絲裂霉素處理的飼養(yǎng)層培養(yǎng)和與少量腹壁組織成纖維細(xì)胞共培養(yǎng)等
[Abstract]:Since Thomson and Gearhart successfully established human embryonic stem cell lines from blastocyst cell population and primordial germ cells in 1998, stem cell research has become the frontier and hot spot in the field of life sciences. The research mainly involves the establishment of embryonic stem cells, inducing stem cells to differentiate into specific tissues and cells, and its application in biology and medicine. Embryonic stem cells (embryonic stem cell,ESC), also known as embryonic stem cells (Embryonic Germ Cell,EGC), derived from primordial germ cells (Primordial Germ Cell,PGC), have the same biological characteristics as embryonic stem cells derived from inner cell group. In this paper, we systematically studied the extraction and expansion of mouse embryonic stem cells, the directional differentiation of embryonic stem cells into hepatocytes, and the experimental study of intrahepatic transplantation of embryonic stem cells. First, the extraction and in vitro expansion of germ stem cells: the extraction and culture of germ stem cells is the prerequisite for in-depth research. We first screened the culture system (see the first part of a series of papers). Like embryonic stem cells derived from inner cell group, embryonic stem cells are easy to differentiate in vitro. In order to maintain the proliferation and undifferentiation of embryonic stem cells in vitro, it is the key to establish an appropriate culture system. Embryonic stem cells require very strict culture conditions in vitro, and different kinds of animal embryonic stem cells require different culture conditions. In recent years, the culture conditions and methods of embryonic stem cells derived from inner cell group have been studied extensively, and abundant experience has been accumulated. However, there are few reports about the culture methods of germ stem cells. In this study, the in vitro culture system suitable for Kunming mouse germ stem cells was screened. The basic culture medium, adding leukemia inhibitory factor, conditioned medium of the same fetal rat fibroblast, feeder layer culture treated with mitomycin and co-culture with a small number of tissue fibroblasts of abdominal wall were observed.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R329

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