本文選題：胚胎干細(xì)胞 + 牙源性上皮�。� 參考：《第四軍醫(yī)大學(xué)》2010年博士論文

【摘要】： 牙齒的發(fā)育過程是依賴于預(yù)定牙齒發(fā)生部位的上皮組織與其下方的間充質(zhì)之間的相互誘導(dǎo)而發(fā)生的。因此,牙齒的生物性再生至少需要兩種細(xì)胞:釉質(zhì)形成所需的牙源性上皮細(xì)胞和形成牙髓牙本質(zhì)復(fù)合體的牙源性間充質(zhì)細(xì)胞。到目前為止,有關(guān)牙源性間充質(zhì)細(xì)胞的替代研究較多,例如成體的牙髓干細(xì)胞、骨髓基質(zhì)細(xì)胞、皮膚的真皮細(xì)胞以及毛囊細(xì)胞等均被證明具有向牙源性間充質(zhì)細(xì)胞轉(zhuǎn)化的能力。然而,在尋找合適的細(xì)胞向牙源性上皮細(xì)胞替代方面成功的研究較少。盡管有學(xué)者認(rèn)為成體骨髓來源的細(xì)胞可以向成釉細(xì)胞轉(zhuǎn)化,但是它可以同時向成釉細(xì)胞樣細(xì)胞和牙源性間充質(zhì)細(xì)胞這兩種細(xì)胞轉(zhuǎn)化的能力限制了它的應(yīng)用。有研究在組織工程牙齒的構(gòu)建中口腔腭粘膜上皮可以替代牙源性上皮,但是此研究必須建立在使用胚胎期或出生1天的自體口腔腭粘膜上皮的基礎(chǔ)上,因此也限制了它的應(yīng)用。這種局限性促使我們尋找一種新的細(xì)胞能夠向牙源性上皮方向分化。胚胎干細(xì)胞具有自我更新和多向分化的能力,近年來受到許多學(xué)者廣泛關(guān)注。有研究證實(shí)胚胎干細(xì)胞可以分化為皮膚表皮細(xì)胞,肺上皮細(xì)胞以及胸腺上皮細(xì)胞等,但是胚胎干細(xì)胞是否可以向牙源性上皮方向轉(zhuǎn)化還尚未見報道。因此對于這一問題的回答,無疑于對解決牙齒再生研究中上皮源性種子細(xì)胞來源的瓶頸問題具有重要意義。本課題主要進(jìn)行了以下幾方面的研究: 1.胚胎干細(xì)胞的培養(yǎng)及鑒定 目的:培養(yǎng)胚胎干細(xì)胞R1系及CGR8系并對其進(jìn)行鑒定。方法:胚胎干細(xì)胞極易受外界因素的影響,對生長條件要求非�？量�。常規(guī)的胚胎干細(xì)胞培養(yǎng)液中含有高糖MEM培養(yǎng)基、胎牛血清、L-谷氨酰胺、2-巰基乙醇和非必需氨基酸等,為了防止胚胎干細(xì)胞發(fā)生分化,還要在培養(yǎng)液中加入白血病抑制因子(LIF),胎牛血清的質(zhì)量及濃度也是影響ES細(xì)胞生長狀況的重要因素,為此本實(shí)驗(yàn)采用GIBCO公司生產(chǎn)的胎牛血清,所加濃度為15%。形成的ES細(xì)胞樣集落,繼續(xù)培養(yǎng)后觀察集落的生長狀態(tài),并通過堿性磷酸酶染色,在體外形成擬胚體,移植入裸鼠皮下4周后取材,組織學(xué)觀察進(jìn)行鑒定。結(jié)果:ES細(xì)胞有其典型的形態(tài)學(xué)特征:集落呈鳥巢狀,邊緣清楚,表面平滑,結(jié)構(gòu)致密,隆起生長,細(xì)胞之間界限不清楚;單個細(xì)胞體積小、核大;對ES細(xì)胞堿性磷酸酶進(jìn)行檢測,在AKP底物作用下,未分化的ES細(xì)胞顯微鏡下為棕褐色,分化的不著色。懸滴法培養(yǎng)ES細(xì)胞兩天后生成擬胚體,它是胚胎干細(xì)胞在體外一定條件下自發(fā)形成的類似早期胚胎的球體結(jié)構(gòu),早期發(fā)育的簡單擬胚體包含了外層的原始內(nèi)胚層和內(nèi)層的原始外胚層,這種結(jié)構(gòu)與胚胎發(fā)育過程中的卵圓柱期結(jié)構(gòu)十分相似。體內(nèi)研究表明,將小鼠胚胎干細(xì)胞注入裸鼠皮下4周后,可形成含有三胚層組織的畸胎瘤。結(jié)論:胚胎干細(xì)胞具有高度的復(fù)制能力及多向分化的潛能。 2.成釉細(xì)胞條件培養(yǎng)液誘導(dǎo)小鼠ES細(xì)胞分化的研究 目的:探討成釉細(xì)胞構(gòu)建的微環(huán)境對于小鼠胚胎干細(xì)胞的誘導(dǎo)能力。方法:本實(shí)驗(yàn)胚胎干細(xì)胞采用兩種途徑分化:生成擬胚體(EB)與不生成擬胚體直接誘導(dǎo)分化的途徑,利用發(fā)育早期成釉細(xì)胞分泌的信號分子在體外模擬成牙微環(huán)境,觀察對小鼠胚胎干細(xì)胞增殖分化的影響。我們用成釉細(xì)胞的條件培養(yǎng)液誘導(dǎo)胚胎干細(xì)胞,并選擇兩個時間點(diǎn),7天和14天,從形態(tài)學(xué)觀察分化后細(xì)胞的形態(tài)特征,并從基因水平和蛋白水平檢測誘導(dǎo)后細(xì)胞的表型變化。隨后我們將兩個誘導(dǎo)組的細(xì)胞植入裸鼠皮下4周觀察體內(nèi)結(jié)果,并將出生1天的C57小鼠的成釉器同樣植入裸鼠皮下作為陽性對照。結(jié)果:體外利用成釉細(xì)胞無血清條件培養(yǎng)液誘導(dǎo)小鼠胚胎干細(xì)胞,形態(tài)學(xué)觀察誘導(dǎo)后可促進(jìn)其增殖和分化,誘導(dǎo)后的部分細(xì)胞形態(tài)與成釉細(xì)胞形態(tài)相似。隨后的基因或蛋白檢測發(fā)現(xiàn)誘導(dǎo)后的細(xì)胞具有牙源性上皮細(xì)胞的表型。其中經(jīng)過EB誘導(dǎo)分化途徑比直接誘導(dǎo)分化途徑的效率要高一些。RT-PCR結(jié)果表明誘導(dǎo)后的胚胎干細(xì)胞表達(dá)牙上皮相關(guān)蛋白CK14、AMBN及AMGN,免疫熒光結(jié)果及Western Blot結(jié)果與RT-PCR結(jié)果一致。體內(nèi)實(shí)驗(yàn)更驗(yàn)證了體外的結(jié)果,將誘導(dǎo)后的小鼠胚胎干細(xì)胞經(jīng)纖維蛋白膠包裹移植于裸鼠皮下,培養(yǎng)4周后觀察經(jīng)過EB誘導(dǎo)分化途徑的細(xì)胞在體內(nèi)可形成一些上皮樣角化組織,進(jìn)一步免疫組織化學(xué)染色顯示角化組織可表達(dá)CK14、AMBN、AMGN,具有類似成釉器樣組織植入裸鼠皮下生成組織的特性。然而不經(jīng)過EB誘導(dǎo)分化途徑的細(xì)胞在體內(nèi)生成了大量的結(jié)締組織,未見典型的上皮樣組織結(jié)構(gòu)。當(dāng)通過擬胚體途徑經(jīng)ASF-CM條件培養(yǎng)液誘導(dǎo)14天的胚胎干細(xì)胞與小鼠牙胚細(xì)胞混合植入裸鼠皮下后,結(jié)果顯示,誘導(dǎo)后的胚胎干細(xì)胞在牙胚細(xì)胞的誘導(dǎo)下,參與了礦化組織的形成,因?yàn)榻?jīng)CFSE標(biāo)記帶綠色熒光的胚胎干細(xì)胞與礦化組織部分重合。未誘導(dǎo)的胚胎干細(xì)胞與牙胚細(xì)胞混合后,體內(nèi)結(jié)果顯示移植物仍然生成畸胎瘤。結(jié)論:以上結(jié)果提示,成釉細(xì)胞無血清條件培養(yǎng)液所提供的成牙微環(huán)境可在體外和體內(nèi)促進(jìn)小鼠胚胎干細(xì)胞的牙向分化,通過擬胚體途徑的分化效率更高一些,且通過擬胚體途徑誘導(dǎo)后的胚胎干細(xì)胞在體內(nèi)能夠生成牙源性上皮樣組織,提示我們胚胎干細(xì)胞向牙源性上皮方向轉(zhuǎn)化的可能性。 3.牙胚細(xì)胞條件培養(yǎng)液誘導(dǎo)小鼠ES細(xì)胞分化的研究 目的:探討牙胚細(xì)胞構(gòu)建的微環(huán)境對于小鼠胚胎干細(xì)胞的誘導(dǎo)能力。方法:本實(shí)驗(yàn)胚胎干細(xì)胞采用兩種途徑分化:生成擬胚體(EB)與不生成擬胚體直接誘導(dǎo)分化的途徑,利用發(fā)育早期牙胚細(xì)胞分泌的信號分子在體外模擬成牙微環(huán)境,觀察對小鼠胚胎干細(xì)胞增殖分化的影響。本實(shí)驗(yàn)主要從形態(tài)學(xué)、基因及蛋白水平進(jìn)行檢測,并將誘導(dǎo)后的細(xì)胞移植入體內(nèi)進(jìn)行觀察。結(jié)果:體外利用牙胚細(xì)胞條件培養(yǎng)液誘導(dǎo)培養(yǎng)小鼠胚胎干細(xì)胞,通過兩種分化途徑,并未實(shí)現(xiàn)向牙源性上皮細(xì)胞方向的表型轉(zhuǎn)化。形態(tài)學(xué)觀察經(jīng)過牙胚條件培養(yǎng)液誘導(dǎo)后,兩種途徑誘導(dǎo)的細(xì)胞增殖能力緩慢,且誘導(dǎo)14天后細(xì)胞出現(xiàn)凋亡�；虮磉_(dá)僅表達(dá)DMP1和CK14,未表達(dá)牙齒上皮樣相關(guān)基因。體內(nèi)移植物為疏松的結(jié)締組織,少量移植物體內(nèi)生成畸胎瘤。未見成釉細(xì)胞及牙齒相關(guān)組織形成。結(jié)論:以上結(jié)果提示,與成釉細(xì)胞條件培養(yǎng)液相比,牙胚細(xì)胞條件培養(yǎng)液所提供的成牙微環(huán)境并不能促進(jìn)小鼠胚胎干細(xì)胞的牙向分化。
[Abstract]:The process of tooth development depends on the mutual induction between the epithelial tissue that is located at the location of the tooth and the mesenchyme at the lower part of the tooth. Therefore, the biological regeneration of the teeth requires at least two kinds of cells: the odontogenic epithelial cells needed for the formation of enamel and the odontogenic mesenchymal cells forming the dental pulp complex. So far, there are many alternative studies on odontogenic mesenchymal cells, such as adult dental pulp stem cells, bone marrow stromal cells, dermal dermal cells and hair follicle cells, which have been proved to have the ability to convert to odontogenic mesenchymal cells. However, the search for the successful substitution of appropriate cells to odontogenic epithelial cells is more successful. Although some scholars believe that the cells derived from adult bone marrow can convert to ameloblastoma, the ability to convert two cells to ameloblastoma like cells and odontogenic mesenchymal cells restricts its application. This study must be based on the use of an embryonic or 1 day autologous oral and palatine epithelium, which also restricts its application. This limitation encourages us to find a new cell to differentiate into the odontogenic epithelium. The ability of embryonic stem cells to be self renewing and pluripotent has received many studies in recent years. Some studies have shown that embryonic stem cells can differentiate into skin epidermal cells, lung epithelial cells and thymic epithelial cells, but it is not yet reported whether embryonic stem cells can convert to odontogenic epithelium. Therefore, the answer to this question is undoubtedly to solve the epithelic seeds in dental regeneration. The bottleneck problem of cell origin is of great significance.
1. culture and identification of embryonic stem cells
Objective: to develop and identify the R1 and CGR8 lines of embryonic stem cells. Methods: embryonic stem cells are very susceptible to external factors and are very demanding for growth conditions. The normal embryonic stem cell culture contains high sugar MEM medium, fetal bovine serum, L- glutamine, 2- mercapto ethanol and non essential amino acids, in order to prevent embryos from embryos. The stem cells are differentiated and the leukemia inhibitory factor (LIF) is added to the culture medium. The quality and concentration of fetal bovine serum are also the important factors affecting the growth of ES cells. Therefore, this experiment adopts the fetal bovine serum produced by GIBCO company, and the concentration is ES fine cell like colony formed by 15%., and the growth state of the colony is observed after continuing culture. With alkaline phosphatase staining, the embryoid body was formed in vitro, and transplanted into the subcutaneous tissue of nude mice for 4 weeks. The results showed that ES cells had its typical morphological characteristics: the colony was nests, the edges were clear, the surface was smooth, the structure was dense, the growth of the cells was not clear, the single cell was small, and the nucleus was large; and ES Cell alkaline phosphatase was detected, under the action of AKP substrate, the undifferentiated ES cell was brown in the microscope, and the differentiation was not coloured. The suspension method was used to produce the ES cells for two days to produce the embryoid body. It was a ball structure similar to the early embryo, which was formed spontaneously under certain conditions in vitro. The original ectoderm of the outer layer of the primordial endoderm and the inner layer of the inner layer, which is very similar to the egg cylindrical structure during the embryonic development, has shown that the mouse embryonic stem cells can form a teratoma containing three germ layers after 4 weeks of subcutaneous injection in nude mice. Conclusion: embryonic stem cells have high replication ability and multidirectional fraction. The potential of transformation.
Differentiation of mouse ES cells induced by 2. ameloblastoma conditioned medium
Objective: To investigate the induction ability of the microenvironment constructed by ameloblastoma to mouse embryonic stem cells. Methods: the experimental embryonic stem cells were divided into two pathways: the generation of EB and the direct induction of differentiation by the non embryoid body, and the observation of the microenvironment by using the early ameloblastoma signal molecules to simulate the microenvironment in vitro. The effect on the proliferation and differentiation of mouse embryonic stem cells. We use the conditioned medium of ameloblastoma to induce embryonic stem cells, and select two time points, 7 days and 14 days, observe the morphological characteristics of the cells after differentiation, and detect the phenotypic changes of the cells after the induction of the gene level and protein level. Then we will take two induction groups. The cells were implanted in the nude mice for 4 weeks to observe the results of the body, and the glaze apparatus of the C57 mice, which was born 1 days, was implanted subcutaneously as a positive control. The morphology of ameloblastoma was similar. Subsequent gene or protein detection found that the induced cells had phenotypes of odontogenic epithelial cells. The EB induced differentiation pathway was more efficient than the direct induction of differentiation pathway..RT-PCR results showed that the induced embryonic stem cells expressed CK14, AMBN and AMGN, and immunofluorescence. The results and the results of Western Blot were in accordance with the results of RT-PCR. In vivo, the results were verified in vitro, and the induced mouse embryonic stem cells were transplanted subcutaneously in nude mice with fibrin glue. After 4 weeks of culture, the cells with EB induced differentiation could form some epitheliated keratinized tissues in the body and further immuno histochemistry The staining showed that the keratinized tissue could express CK14, AMBN, AMGN, which had the characteristics of implanting subcutaneous tissue in nude mice like the enamel like tissue. However, a large number of connective tissues were produced in the body without EB induced differentiation, and no typical epithelioid structure was found. 14 days after the passage of the embryoid pathway through the ASF-CM conditioned medium The results showed that the induced embryonic stem cells were involved in the formation of mineralized tissues under the induction of tooth germ cells, because the CFSE labeled embryonic stem cells with green fluorescence coincided with the mineralized tissue, and the uninduced embryonic stem cells were mixed with the tooth germ cells. The results show that the teratoma is still generated by the graft. Conclusion: the above results suggest that the dental microenvironment provided by the serum-free culture medium of ameloblastoma can promote the tooth differentiation of mouse embryonic stem cells in vitro and in vivo, and the differentiation efficiency through the embryoid pathway is higher, and the embryo fines are induced by the embryoid pathway. Cells can generate odontogenic epithelioid tissue in vivo, suggesting the possibility of embryonic stem cells transferring to the odontogenic epithelium.
Differentiation of mouse ES cells induced by 3. dental germ cell conditioned medium
Objective: To explore the induction ability of the microenvironment constructed by tooth germ cells for mouse embryonic stem cells. Methods: the experimental embryonic stem cells were divided into two ways: the generation of the embryo (EB) and the direct induction of differentiation from the non embryoid body, and using the signal molecules of the early developmental tooth germ cells to simulate the tooth microenvironment in vitro, and observe the microenvironment in vitro. The effect of the experiment on the proliferation and differentiation of mouse embryonic stem cells. This experiment was mainly detected from morphology, gene and protein level, and the induced cells were transplanted into the body for observation. Results: the mouse embryonic stem cells were induced and cultured in vitro by the condition culture solution of tooth germ cell, and the odontogenic epithelium was not finely realized through two differentiation pathways. Phenotypic transformation in the direction of the cell. Morphological observation was induced by the conditioned medium of tooth germ. The cell proliferation ability induced by two pathways was slow and apoptosis was induced in 14 days. Gene expression only expressed DMP1 and CK14 and did not express the epithelioid related genes in the teeth. The body was loose connective tissue and a few transplants were formed to produce malformation. There is no formation of ameloblastoma and dental related tissue. Conclusion: the above results suggest that the dental microenvironment provided by dental germ cell conditioned medium can not promote the tooth differentiation of mouse embryonic stem cells compared with the conditioned medium of ameloblastoma.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2010
【分類號】：R329

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 從玉華;;從皮膚中提取干細(xì)胞[J];中學(xué)生優(yōu)秀作文(中考�？�);2009年10期

2 黃林生;侯玲玲;李向臣;關(guān)偉軍;馬月輝;;牛表皮祖細(xì)胞的體外分離培養(yǎng)[J];安徽農(nóng)業(yè)科學(xué);2011年18期

3 ;國際舞臺上中國干細(xì)胞研究專家2010年4～5月的成果[J];中國組織工程研究與臨床康復(fù);2011年27期

4 朋客;;拋棄男人孕育生命?[J];生命世界;2006年10期

5 徐紅新;田毅浩;曲折;黃文蔚;蔣學(xué)俊;李庚山;;激活Notch對胚胎干細(xì)胞向心肌細(xì)胞分化的影響[J];武漢大學(xué)學(xué)報(醫(yī)學(xué)版);2011年05期

6 郭禮和;;人羊膜上皮細(xì)胞具有胚胎干細(xì)胞和移植免疫耐受等優(yōu)良特性[J];中國細(xì)胞生物學(xué)學(xué)報;2011年04期

7 ;國際舞臺上中國干細(xì)胞研究專家2010年1～2月的學(xué)術(shù)成果[J];中國組織工程研究與臨床康復(fù);2011年23期

8 張樹庸;;科學(xué)家確定DNA的第7種和第8種堿基[J];實(shí)驗(yàn)動物科學(xué);2011年04期

9 ;“人造精子”獲重大突破[J];戀愛婚姻家庭(養(yǎng)生);2011年10期

10 ;國際舞臺上中國干細(xì)胞研究專家2010年6～7月的成果[J];中國組織工程研究與臨床康復(fù);2011年32期

相關(guān)會議論文 前10條

1 秦茂林;蔡文琴;張吉強(qiáng);李成仁;李巍;;雌二醇對胚胎干細(xì)胞神經(jīng)定向分化誘導(dǎo)的影響[A];中國解剖學(xué)會第十一屆全國組織學(xué)與胚胎學(xué)青年學(xué)術(shù)研討會論文匯編[C];2009年

2 徐小明;竇忠英;華進(jìn)聯(lián);葛秀國;;免疫外科法分離克隆BALB/c小鼠胚胎干細(xì)胞[A];全國首屆動物生物技術(shù)學(xué)術(shù)研討會論文集[C];2004年

3 蔣暉;瞿東濱;金大地;鞠躬;;新型神經(jīng)營養(yǎng)因子TAT-BDNF促進(jìn)胚胎干細(xì)胞的神經(jīng)元性分化[A];第一屆全國脊髓損傷治療與康復(fù)研討會暨中國康復(fù)醫(yī)學(xué)會脊柱脊髓損傷專業(yè)委員會脊髓損傷與康復(fù)學(xué)組成立會論文匯編[C];2009年

4 于洲;徐海濱;;應(yīng)用胚胎干細(xì)胞實(shí)驗(yàn)?zāi)Ｐ腕w系對異硫氰酸鹽發(fā)育毒性的研究[A];2010年全國藥物毒理學(xué)學(xué)術(shù)會議論文集[C];2010年

5 劉平;關(guān)云謙;鄒春林;張愚;陳彪;劉焯霖;;胚胎干細(xì)胞在體內(nèi)外誘導(dǎo)分化成為多巴胺能神經(jīng)的初步研究[A];中華醫(yī)學(xué)會第七次全國神經(jīng)病學(xué)學(xué)術(shù)會議論文匯編[C];2004年

6 隋婧;姜方旭;施秉銀;;胚胎干細(xì)胞定向分化為胰島祖細(xì)胞的基因表達(dá)譜分析[A];中華醫(yī)學(xué)會第十次全國內(nèi)分泌學(xué)學(xué)術(shù)會議論文匯編[C];2011年

7 胡敏華;區(qū)穎蕾;李莉;郭欣政;張守全;;胚胎干細(xì)胞飼養(yǎng)層的優(yōu)化及影響內(nèi)細(xì)胞團(tuán)貼壁率的探討[A];中國畜牧獸醫(yī)學(xué)會動物繁殖學(xué)分會第十五屆學(xué)術(shù)研討會論文集（上冊）[C];2010年

8 馬育芳;楊陽;李燕;王少華;趙蕊;李寧;;豬胚胎干細(xì)胞多能性因子Nanog對胚胎發(fā)育的影響[A];中國畜牧獸醫(yī)學(xué)會動物繁殖學(xué)分會第十五屆學(xué)術(shù)研討會論文集（上冊）[C];2010年

9 曹楠;楊黃恬;;大鼠胚胎干細(xì)胞向心肌細(xì)胞體外分化的研究[A];中國病理生理學(xué)會第九屆全國代表大會及學(xué)術(shù)會議論文摘要[C];2010年

10 田孝祥;韓雅玲;康建;徐凱;閆承慧;;利用胚胎干細(xì)胞建立貼壁制備胚胎小體的新方法[A];中華醫(yī)學(xué)會心血管病學(xué)分會第八次全國心血管病學(xué)術(shù)會議匯編[C];2006年

相關(guān)重要報紙文章 前10條

1 張樹庸　趙貴英;干細(xì)胞研究的突破[N];人民日報;2009年

2 記者 鄭曉春;胚胎干細(xì)胞療法安全性研究亟待加強(qiáng)[N];科技日報;2009年

3 記者 余靖靜;我科學(xué)家實(shí)現(xiàn)“用胚胎干細(xì)胞再生肌腱”[N];新華每日電訊;2009年

4 本報記者 李雪林;如何讓中國小實(shí)驗(yàn)室厚積薄發(fā)[N];文匯報;2009年

5 記者孫國根;首個大鼠胚胎干細(xì)胞成功提取[N];健康報;2009年

6 孫國根;復(fù)旦大學(xué)成功提取首個大鼠胚胎干細(xì)胞[N];中國醫(yī)藥報;2009年

7 本報首席記者 任荃;第二顆“全能種子”會不會搶先“發(fā)芽”[N];文匯報;2009年

8 記者 鄭曉春;懸浮液可用于大規(guī)模培育胚胎干細(xì)胞[N];科技日報;2010年

9 記者 錢錚;牛膠原蛋白可助降低胚胎干細(xì)胞癌變幾率[N];新華每日電訊;2009年

10 記者 任海軍;奧兌現(xiàn)承諾,但“松綁”效應(yīng)不會立竿見影[N];新華每日電訊;2009年

相關(guān)博士學(xué)位論文 前10條

1 譚舟;MK和GM-CSFRα在胚胎干細(xì)胞中的表達(dá)與功能研究[D];浙江大學(xué);2010年

2 寧芳;誘導(dǎo)小鼠胚胎干細(xì)胞向牙齒上皮樣細(xì)胞分化的實(shí)驗(yàn)研究[D];第四軍醫(yī)大學(xué);2010年

3 蘇中淵;胚胎干細(xì)胞來源的間充質(zhì)干細(xì)胞歸巢及胚胎干細(xì)胞表面分子的研究[D];浙江大學(xué);2010年

4 廖宏慶;人類不同原核數(shù)目受精卵中篩選正常優(yōu)質(zhì)胚胎的策略研究[D];中南大學(xué);2010年

5 白春玲;牛孤雌多倍體胚胎與克隆多倍體胚胎的研究[D];內(nèi)蒙古大學(xué);2011年

6 曲文玉;人胚胎干細(xì)胞的分離、培養(yǎng)及建系[D];中國醫(yī)科大學(xué);2006年

7 李相運(yùn);小鼠胚胎干細(xì)胞多能性研究[D];西北農(nóng)林科技大學(xué);2001年

8 姚嘉宜;胚胎干細(xì)胞重新編程腫瘤細(xì)胞的研究[D];第二軍醫(yī)大學(xué);2008年

9 趙惠萍;骨髓內(nèi)皮細(xì)胞條件培養(yǎng)液誘導(dǎo)胚胎干細(xì)胞向造血細(xì)胞分化[D];中南大學(xué);2003年

10 劉星霞;胚胎干細(xì)胞ES-D_3誘導(dǎo)分化為胰島素分泌細(xì)胞及其對糖尿病鼠降糖作用的研究[D];山東大學(xué);2005年

相關(guān)碩士學(xué)位論文 前10條

1 孫國杰;山羊類胚胎干細(xì)胞的分離、克隆[D];河北農(nóng)業(yè)大學(xué);2002年

2 呂一帆;胚胎干細(xì)胞在內(nèi)膜損傷小鼠宮腔內(nèi)移植的實(shí)驗(yàn)研究[D];福建醫(yī)科大學(xué);2010年

3 周文平;小鼠三種胚胎干細(xì)胞表觀相關(guān)基因的分析[D];河南大學(xué);2011年

4 姜靜宜;胚胎干細(xì)胞中外源性IbeB靶蛋白的篩選及驗(yàn)證[D];中國醫(yī)科大學(xué);2010年

5 蔡斌;體外誘導(dǎo)胚胎干細(xì)胞為神經(jīng)細(xì)胞模型的建立及相關(guān)因子的實(shí)驗(yàn)研究[D];第二軍醫(yī)大學(xué);2001年

6 劉小春;轉(zhuǎn)化生長因子β_1聯(lián)合血管內(nèi)皮細(xì)胞生長因子體外誘導(dǎo)胚胎干細(xì)胞向內(nèi)皮細(xì)胞分化[D];南昌大學(xué);2007年

7 劉艷;山羊類胚胎干細(xì)胞的分離培養(yǎng)及其來源的上皮樣細(xì)胞的培養(yǎng)[D];華中農(nóng)業(yè)大學(xué);2010年

8 顧文佳;誘導(dǎo)胚胎干細(xì)胞向神經(jīng)細(xì)胞分化的研究[D];中國醫(yī)科大學(xué);2004年

9 劉愿;小鼠球形精子受精胚胎制備及培養(yǎng)方法的改進(jìn)[D];山東師范大學(xué);2011年

10 秦茂林;小鼠胚胎干細(xì)胞的分離、鑒定和神經(jīng)定向誘導(dǎo)分化[D];第三軍醫(yī)大學(xué);2001年

，

本文編號：2074398