缺血損傷時(shí)骨髓間充質(zhì)干細(xì)胞在種間歸巢能力的實(shí)驗(yàn)研究
發(fā)布時(shí)間：2018-11-14 14:02
【摘要】：目的：研究缺血損傷時(shí)骨髓間充質(zhì)干細(xì)胞(bone Mesenchymal stem cells, BMSCs)在種間的歸巢能力。 方法：1.EGFP-BMSCs|的分離培養(yǎng)及純度鑒定：貼壁培養(yǎng)法擴(kuò)增增強(qiáng)型綠色熒光蛋白(enhanced green fluorescent protein, EGFP)轉(zhuǎn)基因C57BL/6小鼠的EGFP-BMSCs,體外傳代至第5代,并行流式細(xì)胞學(xué)鑒定EGFP-BMSCs的純度。 2.缺血損傷模型的建立：受鼠麻醉后,雙下肢大腿外側(cè)剪毛消毒,右側(cè)股部經(jīng)前外側(cè)切口切開(kāi),顯露、分離股四頭肌并用1號(hào)絲線(xiàn)結(jié)扎近、遠(yuǎn)端,止血關(guān)閉切口。左側(cè)切開(kāi)皮膚,股四頭肌不分離不結(jié)扎,作為對(duì)照,止血后關(guān)閉切口。其他小鼠同法建立缺血模型,大鼠缺血模型分離股四頭肌外側(cè)頭結(jié)扎。 3.消化相同供鼠(EGFP轉(zhuǎn)基因小鼠)的BMSCs,計(jì)數(shù)并調(diào)整濃度為2×106個(gè)/ml,取0.1ml經(jīng)尾靜脈注入受鼠。分別于第1天、4天、7天、14天、28天處死受鼠,取右側(cè)缺血肌肉模型及對(duì)側(cè)肌肉組織,提取總蛋白,進(jìn)行Western blotting檢測(cè)EGFP,掃描并做灰度分析,統(tǒng)計(jì)分析。 結(jié)果：1.經(jīng)Western blotting檢測(cè)EGFP,缺血側(cè)有EGFP表達(dá),說(shuō)明異體EGFP-BMSCs植入受鼠后,EGFP-BMSCs可歸巢到缺血組織。 2.經(jīng)統(tǒng)計(jì)分析各時(shí)間段種間缺血側(cè)的灰度值無(wú)差異,P0.05,無(wú)統(tǒng)計(jì)學(xué)意義,表明種間EGFP-BMSCs歸巢沒(méi)有差異,異種EGFP-BMSCs具有低免疫原性。 結(jié)論：1BMSCs可以歸巢到損傷組織。 2.利用EGFP轉(zhuǎn)基因小鼠的BMSCs可以很好的示蹤BMSCs的歸巢。 3. BMSCs種間歸巢沒(méi)有差異,BMSCs具有低免疫原性。 4. BMSCs歸巢是一種以主動(dòng)運(yùn)動(dòng)為主的遷移。
[Abstract]:Aim: to study the homing ability of bone marrow mesenchymal stem cells (bone Mesenchymal stem cells, BMSCs) during ischemic injury. Methods: isolation, culture and purity identification of 1.EGFP-BMSCs: the EGFP-BMSCs, of (enhanced green fluorescent protein, EGFP) transgenic C57BL/6 mice with enhanced green fluorescent protein (GFP) was amplified by adherent culture method and was subcultured to the 5th generation in vitro. The purity of EGFP-BMSCs was identified by flow cytology. 2. The establishment of ischemic injury model: after anaesthesia, the lateral thigh shearing of both lower limbs was disinfected, the right thigh was cut and exposed through the anterolateral incision, the quadriceps femoris was separated and ligated, the distal side was ligated and the incision was closed by hemostasis. Left incision skin, quadriceps femoris not separated and not ligated, as a control, hemostasis closed incision. The ischemia model was established by the same method in other mice, and the lateral head of quadriceps femoris was isolated from the ischemic model of rats. 3. The BMSCs, count of the same donor mice (EGFP transgenic mice) was digested and the concentration of 0.1ml was adjusted to 2 脳 106 / ml,. 0.1ml was injected into recipient mice via tail vein. The recipient rats were killed on day 1, day 4, day 7, day 14 and day 28, respectively. The right ischemic muscle model and contralateral muscle tissue were taken, total protein was extracted, EGFP, scan was performed by Western blotting and gray scale analysis was performed. Results: 1. The expression of EGFP in ischemic side of EGFP, was detected by Western blotting, indicating that EGFP-BMSCs could homing to ischemic tissue after allogeneic EGFP-BMSCs implantation. 2. Statistical analysis showed that there was no difference in grayscale value of ischemic side between different species and there was no significant difference in homing of EGFP-BMSCs between species and in different species EGFP-BMSCs was low immunogenicity. Conclusion: 1BMSCs can homing to damaged tissue. 2. The BMSCs of EGFP transgenic mice can well trace the homing of BMSCs. 3. There was no difference in homing among BMSCs species, and BMSCs had low immunogenicity. 4. BMSCs homing is an active migration.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R329
本文編號(hào)：2331363
[Abstract]:Aim: to study the homing ability of bone marrow mesenchymal stem cells (bone Mesenchymal stem cells, BMSCs) during ischemic injury. Methods: isolation, culture and purity identification of 1.EGFP-BMSCs: the EGFP-BMSCs, of (enhanced green fluorescent protein, EGFP) transgenic C57BL/6 mice with enhanced green fluorescent protein (GFP) was amplified by adherent culture method and was subcultured to the 5th generation in vitro. The purity of EGFP-BMSCs was identified by flow cytology. 2. The establishment of ischemic injury model: after anaesthesia, the lateral thigh shearing of both lower limbs was disinfected, the right thigh was cut and exposed through the anterolateral incision, the quadriceps femoris was separated and ligated, the distal side was ligated and the incision was closed by hemostasis. Left incision skin, quadriceps femoris not separated and not ligated, as a control, hemostasis closed incision. The ischemia model was established by the same method in other mice, and the lateral head of quadriceps femoris was isolated from the ischemic model of rats. 3. The BMSCs, count of the same donor mice (EGFP transgenic mice) was digested and the concentration of 0.1ml was adjusted to 2 脳 106 / ml,. 0.1ml was injected into recipient mice via tail vein. The recipient rats were killed on day 1, day 4, day 7, day 14 and day 28, respectively. The right ischemic muscle model and contralateral muscle tissue were taken, total protein was extracted, EGFP, scan was performed by Western blotting and gray scale analysis was performed. Results: 1. The expression of EGFP in ischemic side of EGFP, was detected by Western blotting, indicating that EGFP-BMSCs could homing to ischemic tissue after allogeneic EGFP-BMSCs implantation. 2. Statistical analysis showed that there was no difference in grayscale value of ischemic side between different species and there was no significant difference in homing of EGFP-BMSCs between species and in different species EGFP-BMSCs was low immunogenicity. Conclusion: 1BMSCs can homing to damaged tissue. 2. The BMSCs of EGFP transgenic mice can well trace the homing of BMSCs. 3. There was no difference in homing among BMSCs species, and BMSCs had low immunogenicity. 4. BMSCs homing is an active migration.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R329
【參考文獻(xiàn)】
相關(guān)期刊論文 前6條
1 向國(guó)安,張剛慶,方馳華,高鵬,陳開(kāi)運(yùn);同種異體骨髓間質(zhì)干細(xì)胞移植在大鼠肝內(nèi)定居能力初步研究[J];第一軍醫(yī)大學(xué)學(xué)報(bào);2005年08期
2 田瑩;鄧宇斌;王亞柱;王曄;朱文標(biāo);;骨髓間質(zhì)干細(xì)胞降低大鼠移植物抗宿主反應(yīng)的實(shí)驗(yàn)研究[J];免疫學(xué)雜志;2008年01期
3 王濤;李露斯;劉雁平;屈紀(jì)富;徐海偉;;脊髓損傷后真皮多能干細(xì)胞移植時(shí)機(jī)的選擇[J];中國(guó)組織工程研究與臨床康復(fù);2007年33期
4 施小鳳,傅晉翔,梁建英,張宏,張勇,張曉慧;骨髓間充質(zhì)干細(xì)胞體內(nèi)歸巢特性的研究[J];江蘇醫(yī)藥;2004年04期
5 周進(jìn)明,鄒仲敏,郭朝華,何穎,朱波,王蒙,冉新澤,羅成基,程天民;培養(yǎng)小鼠骨髓間充質(zhì)干細(xì)胞及其移植后在體內(nèi)的定位分布[J];中華放射醫(yī)學(xué)與防護(hù)雜志;2002年03期
6 楊光;范東艷;陳強(qiáng);任淑萍;尹維田;;貼壁法培養(yǎng)不同齡小鼠骨髓間充質(zhì)干細(xì)胞的生物學(xué)特點(diǎn)[J];中國(guó)實(shí)驗(yàn)診斷學(xué);2007年01期
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