【摘要】：ROSAβgeo26(ROSA26)小鼠線系是由胚胎干細(xì)胞線系衍生而來(lái)，其在胚胎發(fā)育過(guò)程中的所有細(xì)胞均表達(dá)報(bào)告基因LacZ，故被視為骨髓移植實(shí)驗(yàn)中的一個(gè)有價(jià)值的標(biāo)記物線系。LacZ基因的產(chǎn)物β-半乳糖苷酶在ROSA26小鼠線系的全身組織廣泛表達(dá)。然而，，β-半乳糖苷酶在該小鼠線系腦中的細(xì)胞分布和轉(zhuǎn)基因神經(jīng)干細(xì)胞在移植中的利用還有待于研究。為此，本實(shí)驗(yàn)采用X-Gal染色、尼氏染色和免疫組織及細(xì)胞化學(xué)染色方法研究了β-半乳糖苷酶在ROSA26小鼠的腦和神經(jīng)干細(xì)胞中的表達(dá)特征。結(jié)果顯示：β-半乳糖苷酶在新生和成年ROSA26小鼠腦的神經(jīng)干細(xì)胞和神經(jīng)元中廣泛表達(dá)，而在星形膠質(zhì)細(xì)胞中的表達(dá)下調(diào)。轉(zhuǎn)基因神經(jīng)干細(xì)胞和星形膠質(zhì)細(xì)胞的培養(yǎng)實(shí)驗(yàn)進(jìn)一步證實(shí)了在體實(shí)驗(yàn)的研究結(jié)果。移植實(shí)驗(yàn)顯示整合到宿主大鼠腦中的轉(zhuǎn)基因神經(jīng)干細(xì)胞在分化后的神經(jīng)元具有β-半乳糖苷酶活性，而星形膠質(zhì)細(xì)胞則不表達(dá)β-半乳糖苷酶的活性。上述結(jié)果提示ROSA26小鼠中的LacZ基因在神經(jīng)干細(xì)胞分化為膠質(zhì)細(xì)胞系的過(guò)程中沉默，故在移植研究中評(píng)價(jià)轉(zhuǎn)基因細(xì)胞在宿主體內(nèi)命運(yùn)時(shí)應(yīng)注意此現(xiàn)象。
[Abstract]:ROSA 尾 geo26 (ROSA26) mouse line is derived from embryonic stem cell line. The reporter gene LacZ, was expressed in all the cells during embryonic development, so it was regarded as a valuable marker line of bone marrow transplantation. The product of Lacz gene 尾 -galactosidase was widely expressed in the whole body of ROSA26 mice. However, the cell distribution of 尾 -galactosidase in the mouse linear brain and the use of transgenic neural stem cells in transplantation need to be studied. Therefore, the expression of 尾 -galactosidase in the brain and neural stem cells of ROSA26 mice was studied by X-Gal staining, Nissl staining, immunohistochemistry and cytochemical staining. The results showed that 尾 -galactosidase was widely expressed in neural stem cells and neurons of newborn and adult ROSA26 mice, but down-regulated in astrocytes. Transgenic neural stem cells and astrocytes culture experiments further confirmed the results of in vivo experiments. Transplantation experiments showed that the differentiated neurons of transgenic neural stem cells integrated into host rat brain had 尾 -galactosidase activity, while astrocytes did not express 尾 -galactosidase activity. These results suggest that the LacZ gene in ROSA26 mice is silenced during the differentiation of neural stem cells into glial cell lines, so we should pay attention to this phenomenon when evaluating the fate of transgenic cells in vivo.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 邢文英;金輝;丁一;;衰老相關(guān)-β-半乳糖苷酶和P16在老年大鼠脾內(nèi)的表達(dá)及意義[J];解剖學(xué)雜志;2008年04期

2 周國(guó)棟;田耕;畢伏龍;;神經(jīng)干細(xì)胞移植的研究近況[J];內(nèi)蒙古民族大學(xué)學(xué)報(bào)(自然科學(xué)版);2006年02期

3 王娜;劉硯星;連易水;熊晨;;神經(jīng)干細(xì)胞研究進(jìn)展[J];生物學(xué)雜志;2009年01期

4 ;Experimental Study of Cell Migration and Functional Differentiation of Transplanted Neural Stem Cells Co-labeled with Superparamagnetic Iron Oxide and Brdu in an Ischemic Rat Model[J];Biomedical and Environmental Sciences;2008年05期

5 楊林;夏鷹;趙洪洋;趙甲山;朱賢立;;Magnetic Resonance Imaging of Transplanted Neural Stem Cells in Parkinson Disease Rats[J];華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)英德文版);2006年04期

6 楮倩;喻志源;張?zhí)K明;于世英;;Astrocytes Facilitate the Growth and Differentiation of Co-cultured Mesenchymal Stem Cells[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2008年03期

7 翟寶進(jìn);陳禮明;;神經(jīng)干細(xì)胞應(yīng)用研究進(jìn)展[J];武警醫(yī)學(xué)院學(xué)報(bào);2008年11期

8 楊玉芳;轉(zhuǎn)基因小鼠技術(shù)在腫瘤研究中的應(yīng)用與進(jìn)展[J];醫(yī)學(xué)綜述;2004年01期

本文編號(hào)：2211172