本文選題：人羊膜上皮細(xì)胞 + 誘導(dǎo)分化��； 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：背景:近年來(lái),關(guān)于干細(xì)胞治療的研究越來(lái)越火熱,而在臨床治療的應(yīng)用上,對(duì)眾多干細(xì)胞的選擇是其關(guān)鍵問(wèn)題。人羊膜組織作為生產(chǎn)的附屬品,來(lái)源豐富。人羊膜來(lái)源的干細(xì)胞,尤其是人羊膜上皮細(xì)胞(human Amnion Epithelial Cells,hAECs)具有向三胚層分化的潛能,表達(dá)多種胚胎干細(xì)胞標(biāo)記物,并且hAECs具有無(wú)致瘤性、來(lái)源豐富、容易獲取、免疫原性低、無(wú)醫(yī)學(xué)倫理限制等優(yōu)點(diǎn),很多優(yōu)勢(shì)是胚胎干細(xì)胞和諸多成體干細(xì)胞所不具備的。肝臟是人體重要的代謝、排毒、免疫器官,對(duì)人體至關(guān)重要,但嚴(yán)重的肝臟疾病目前還沒(méi)有可以完全治愈的治療措施。因此hAECs有望成為今后臨床移植治療的重要種子細(xì)胞。方法:使用酶消化法從人羊膜中分離得到hAECs,觀察細(xì)胞形態(tài),并繪制第2代和第5代hAECs的生長(zhǎng)曲線。免疫細(xì)胞化學(xué)法檢測(cè)hAECs角蛋白19的表達(dá)。RT-PCR鑒定hAECs表達(dá)干細(xì)胞相關(guān)基因。用四氯化碳(CC14)腹腔注射誘導(dǎo)小鼠肝損傷。將熒光染料羥基熒光素二醋酸鹽琥珀酰亞胺脂(CFSE)標(biāo)記的hAECs通過(guò)尾靜脈注射移植到肝損傷小鼠體內(nèi),檢測(cè)細(xì)胞移植后小鼠肝功、組織病理學(xué)變化。結(jié)果:hAECs核大、形態(tài)呈鋪路石狀,可在體外連續(xù)傳代培養(yǎng)。免疫細(xì)胞化學(xué)檢測(cè)顯示hAECs表達(dá)上皮細(xì)胞標(biāo)志角蛋白19。RT-PCR檢測(cè)顯示hAECs表達(dá)干細(xì)胞標(biāo)志基因REX1、Oct-4和Nanog。體外可成功誘導(dǎo)為脂肪細(xì)胞、神經(jīng)細(xì)胞和肝樣細(xì)胞,并在誘導(dǎo)后表達(dá)相應(yīng)細(xì)胞的標(biāo)志基因PPARγ2、MAP-2和albumin。冰凍切片免疫熒光顯示hAECs可以歸巢到受損肝臟。hAECs移植后對(duì)肝損傷小鼠的肝功、組織病理學(xué)都有修復(fù)作用,治療效果顯著。結(jié)論:人羊膜上皮細(xì)胞可以用于異種移植。移植到四氯化碳誘導(dǎo)肝損傷模型小鼠體內(nèi),對(duì)于損傷肝臟恢復(fù)具有促進(jìn)作用。
[Abstract]:Background: in recent years, the research on stem cell therapy has become more and more hot, and the selection of stem cells is the key issue in clinical treatment. Human amniotic tissue, as a subsidiary of production, is rich in sources. Stem cells derived from human amniotic membrane, especially human amniotic epithelial cells (HMECs), have the potential to differentiate into the tridermal layer and express a variety of embryonic stem cell markers. Moreover, hAECs has non-tumorigenic properties, rich sources, easy to obtain, and low immunogenicity. Without medical ethics restrictions, many advantages are not available to embryonic stem cells and adult stem cells. Liver is an important metabolism, detoxification and immune organ, which is very important to human body. However, there is no cure for serious liver diseases. Therefore, hAECs is expected to be an important seed cell for clinical transplantation in the future. Methods: hAECs were isolated from human amniotic membrane by enzyme digestion. The morphology of the cells was observed and the growth curves of the second and fifth generation hAECs were plotted. The expression of hAECs keratin 19 was detected by immunocytochemistry. RT-PCR was used to identify the stem cell related genes expressed by hAECs. CC14) was used to induce liver injury in mice. HAECs labeled with fluorescein hydroxy fluorescein diacetate succinimide (CFSE) was transplanted into mice with liver injury by tail vein injection. The liver function and histopathological changes were detected after transplantation. Results the nucleus of 1% hAECs was large and its morphology was paving stone, which could be subcultured continuously in vitro. Immunocytochemistry showed that hAECs expressed epithelial cell marker keratin 19.RT-PCR and hAECs expressed stem cell marker REX1 Oct-4 and Nanog4. Adipocytes, nerve cells and hepatoid cells were successfully induced in vitro, and the marker genes PPAR 緯 2 MAP-2 and Albumin were expressed after induction. The immunofluorescence of frozen sections showed that hAECs could homing to the injured liver. After transplantation, the liver function and histopathology of the injured mice were repaired, and the therapeutic effect was remarkable. Conclusion: human amniotic epithelial cells can be used in xenotransplantation. Transplantation into carbon tetrachloride induced liver injury model mice can promote the recovery of injured liver.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R575

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