【摘要】：ROSAβgeo26(ROSA26)小鼠線系是由胚胎干細胞線系衍生而來，其在胚胎發(fā)育過程中的所有細胞均表達報告基因LacZ，故被視為骨髓移植實驗中的一個有價值的標記物線系。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。然而，，β-半乳糖苷酶在該小鼠線系腦中的細胞分布和轉基因神經(jīng)干細胞在移植中的利用還有待于研究。為此，本實驗采用X-Gal染色、尼氏染色和免疫組織及細胞化學染色方法研究了β-半乳糖苷酶在ROSA26小鼠的腦和神經(jīng)干細胞中的表達特征。結果顯示：β-半乳糖苷酶在新生和成年ROSA26小鼠腦的神經(jīng)干細胞和神經(jīng)元中廣泛表達，而在星形膠質(zhì)細胞中的表達下調(diào)。轉基因神經(jīng)干細胞和星形膠質(zhì)細胞的培養(yǎng)實驗進一步證實了在體實驗的研究結果。移植實驗顯示整合到宿主大鼠腦中的轉基因神經(jīng)干細胞在分化后的神經(jīng)元具有β-半乳糖苷酶活性，而星形膠質(zhì)細胞則不表達β-半乳糖苷酶的活性。上述結果提示ROSA26小鼠中的LacZ基因在神經(jīng)干細胞分化為膠質(zhì)細胞系的過程中沉默，故在移植研究中評價轉基因細胞在宿主體內(nèi)命運時應注意此現(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.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R329

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