發(fā)布時(shí)間：2018-01-31 04:39

  本文關(guān)鍵詞： 腦創(chuàng)傷 骨髓間充質(zhì)干細(xì)胞 超順磁性氧化鐵顆粒 示蹤 磁共振成像　出處：《福建醫(yī)科大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 分離、培養(yǎng)、鑒定及SPIO標(biāo)記大鼠BMSCs,探討MRI對其在大鼠腦創(chuàng)傷模型活體示蹤成像中的可行性。 方法 1.全骨髓貼壁培養(yǎng)法分離培養(yǎng)大鼠BMSCs,流式細(xì)胞儀檢測細(xì)胞表面抗原及向成脂、成脂誘導(dǎo)分化進(jìn)行鑒定,并探討首次換液模式對BMSCs培養(yǎng)時(shí)間及細(xì)胞純度的影響。 2.以左旋多聚賴氨酸(PLL)為轉(zhuǎn)染介質(zhì)介導(dǎo)導(dǎo)BMSCs吞噬SPIO,評價(jià)SPIO對細(xì)胞生物學(xué)特性的影響,探討最佳SPIO濃度。 3.參照Feeney法制作大鼠TBI模型,經(jīng)右側(cè)經(jīng)總動脈穿刺插管注射移植SPIO標(biāo)記的BMSCs。 4.實(shí)驗(yàn)組大鼠注射移植SPIO標(biāo)記BMSCs,對照組TBI大鼠注射等量的無SPIO標(biāo)記的BMSCs或PBS,均分別于注射細(xì)胞前及注射BMSCs后1d、3d、7d、14d、21d進(jìn)行NSS評分及MRI掃描,觀察腦實(shí)質(zhì)內(nèi)信號變化及其遷移情況,并與對側(cè)正常腦實(shí)質(zhì)信號進(jìn)行對比。每次MRI掃描結(jié)束后立即各處死1只,取腦組織切片進(jìn)行HE及普魯士藍(lán)染色。 結(jié)果 1.BMSCs生長狀態(tài)良好,培養(yǎng)時(shí)間較短,純度較高,CD29、CD90陽性,CD34、CD45陰性,向成骨、成脂誘導(dǎo)分化,SOIO標(biāo)記后細(xì)胞生物活性良好。 2.MRI掃描可觀察到實(shí)驗(yàn)組TBI模型大鼠腦實(shí)質(zhì)內(nèi)低信號區(qū),并逐步向病變區(qū)遷移,與對側(cè)腦實(shí)質(zhì)內(nèi)信號減低率結(jié)果具有統(tǒng)計(jì)學(xué)意義(F=154.19,P0.05),病理切片普魯士藍(lán)染色見藍(lán)染顆粒。BMSCs移植組NSS評分低于PBS移植組,結(jié)果有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論 1.全骨髓貼壁法培養(yǎng)BMSCs生物學(xué)特性良好,可被SPIO有效標(biāo)記且不影響生物活性。 2. 3.0T MRI聯(lián)合動物專用線圈對SPIO標(biāo)記的BMSCs活體示蹤有意義。 3.BMSCs可能對大鼠神經(jīng)功能恢復(fù)有一定作用。
[Abstract]:Purpose To investigate the feasibility of MRI in the in vivo tracer imaging of rat brain trauma model, we isolated, cultured, identified and labeled BMSCs with SPIO. Method 1. Whole bone marrow adherent culture method was used to isolate and culture BMSCs. Flow cytometry was used to detect cell surface antigen and adipogenic differentiation. The effects of the first liquid exchange model on the culture time and cell purity of BMSCs were investigated. 2. The phagocytosis of BMSCs mediated by L-polylysine (PLL) was studied to evaluate the effect of SPIO on cell biological characteristics and to explore the optimal concentration of SPIO. 3. The rat model of TBI was made by referring to the method of Feeney, and the BMSCs labeled by SPIO were injected into the right common artery via the right side. 4. The experimental group was injected with SPIO labeled BMSCs, while the control group with TBI was injected with the same amount of BMSCs or PBS without SPIO labeling. NSS scores and MRI scans were performed before the injection of the cells and 1 day after BMSCs injection. The changes of signal and migration in the brain parenchyma were observed. At the end of each MRI scan, 1 rat was killed and the brain sections were stained with HE and Prussian blue. Results 1. BMSCs grow well, culture time is short, the purity of BMSCs is high, CD29 + CD90 + CD34 + CD34 CD45 negative, osteogenesis, adipogenic differentiation. SOIO labeled cells showed good bioactivity. 2. The hypointense area in the brain parenchyma of the experimental group TBI model rats was observed by MRI scanning, and the brain moved to the lesion area gradually. The results of signal reduction in the contralateral cerebral parenchyma were statistically significant (P 0.05). Prussian blue staining of pathological sections showed that the NSS score of blue dye granules. BMSCs transplantation group was lower than that of PBS transplantation group, and the result was statistically significant (P 0.05). Conclusion 1. The biological characteristics of BMSCs cultured by whole bone marrow adherent method were good, which could be effectively labeled by SPIO and had no effect on biological activity. 2.3.0T MRI combined with animal coil has significance for SPIO labeled BMSCs tracer in vivo. 3. BMSCs may play a role in the recovery of neural function in rats.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R-332

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