發(fā)布時間：2019-01-27 21:57

【摘要】：肝臟作為機(jī)體最重要的生命器官之一,在機(jī)體的代謝、排毒、循環(huán)調(diào)節(jié)等方面發(fā)揮著重要的功能,同時肝臟疾病也是影響人體健康的重要疾病之一,目前,對于一些嚴(yán)重的末期肝臟疾病,諸如肝癌,急性肝衰竭等,最為有效的治療手段就是原位肝移植,然而由于供體肝缺乏,高昂的費用,以及異種免疫排斥等原因,使其臨床應(yīng)用受到很大的限制。因此,一種新的以細(xì)胞為基礎(chǔ)的肝病治療法,例如肝組織工程,旨在造出一個人造肝組織使其成為患者肝組織的替代品。肝臟組織工程為肝病的治療帶來了新的希望。 近年來間充質(zhì)干細(xì)胞的研究和應(yīng)用備受關(guān)注,間充質(zhì)干細(xì)胞(mesenchymal stem cells, MSCs)具有很強增殖能力和多分化潛能,具有跨胚層分化能力,而且間充質(zhì)干細(xì)胞容易獲得,易于分離培養(yǎng)和擴(kuò)增。因此間充質(zhì)干細(xì)胞(MSCs)作為組織工程的種子細(xì)胞具有良好的應(yīng)用前景。在本論文中間充質(zhì)干細(xì)胞分離自人臍帶沃頓膠質(zhì)組織(Wharton’s jelly)經(jīng)過體外誘導(dǎo)培養(yǎng)分化為肝樣細(xì)胞,并將其與骨髓來源的間充質(zhì)干細(xì)胞(BM-MSCs)進(jìn)行比較。研究表明臍帶間充質(zhì)干細(xì)胞和骨髓間充質(zhì)干細(xì)胞都具有相似的間充質(zhì)干細(xì)胞特性,都能在體外誘導(dǎo)分化為成骨細(xì)胞,脂肪細(xì)胞和軟骨細(xì)胞。經(jīng)過肝細(xì)胞誘導(dǎo)分化液的誘導(dǎo)培養(yǎng)后,分化后的細(xì)胞都能夠表達(dá)肝細(xì)胞特異性的蛋白和基因(albumin, AFP, CK-18)并且具有糖原合成能力和低密度脂蛋白吸收能力,結(jié)果表明臍帶間充質(zhì)干細(xì)胞和骨髓間充質(zhì)干細(xì)胞具有相似的肝細(xì)胞分化潛能。但是與骨髓間充質(zhì)干細(xì)胞相比,臍帶間充質(zhì)干細(xì)胞更容易獲得,具有低的免疫原性,所以臍帶間充質(zhì)干細(xì)胞作為組織工程的種子細(xì)胞更具優(yōu)勢。聚羥基脂肪酸酯(PHA)是微生物在營養(yǎng)不平衡的環(huán)境下產(chǎn)生的一類高分子生物線性聚酯,研究表明PHA有良好的生物相容性及生物可降解性,三聚物PHBVHHx與其他的PHA材料相比具有更好的生物相容性,更利于細(xì)胞的貼附生長,所以PHBVHHx在肝組織工程的研究上具有很好的應(yīng)用前景。 本研究中我們選擇臍帶間充質(zhì)干細(xì)胞作為肝組織工程的種子細(xì)胞,三聚物PHBVHHx作為支架材料,研究生物材料PHBVHHx支架對臍帶間充質(zhì)干細(xì)胞體外分化為肝細(xì)胞的影響。本實驗研究結(jié)果表明PHBVHHx支架能夠支持臍帶間充質(zhì)干細(xì)胞的生長,并且臍帶間充質(zhì)干細(xì)胞在PHBVHHx支架上在肝細(xì)胞誘導(dǎo)液的誘導(dǎo)培養(yǎng)下能夠分化肝樣細(xì)胞。總之本研究可以為肝組織工程的研究提供一定的實驗信息和數(shù)據(jù)。
[Abstract]:As one of the most important living organs of the body, liver plays an important role in metabolism, detoxification, circulatory regulation and so on. At the same time, liver disease is also one of the important diseases affecting human health. For some serious end-stage liver diseases, such as liver cancer and acute liver failure, the most effective treatment is orthotopic liver transplantation. However, due to lack of donor liver, high cost, and xenoimmune rejection, etc. Its clinical application is restricted greatly. Therefore, a new cell-based treatment for liver diseases, such as liver tissue engineering, aims to create an artificial liver tissue as a substitute for patient liver tissue. Liver tissue engineering offers new hope for the treatment of liver disease. In recent years, the research and application of mesenchymal stem cells (MSCs) have attracted much attention. Mesenchymal stem cells (mesenchymal stem cells, MSCs) have strong proliferative ability, multi-differentiation potential, transdermal differentiation ability and easy to obtain mesenchymal stem cells. It is easy to isolate, culture and amplify. Therefore, mesenchymal stem cell (MSCs) as seed cells of tissue engineering has a good application prospect. Mesenchymal stem cells (MSCs) isolated from human umbilical cord Wharton glial tissue (Wharton's jelly) were induced to differentiate into hepatoid cells in vitro and were compared with bone marrow-derived mesenchymal stem cells (BM-MSCs). The results showed that both umbilical cord mesenchymal stem cells and bone marrow mesenchymal stem cells had similar characteristics of mesenchymal stem cells and could be induced to differentiate into osteoblasts, adipocytes and chondrocytes in vitro. The differentiated cells were able to express hepatocyte specific protein and gene (albumin, AFP, CK-18) and had the ability of glycogen synthesis and low density lipoprotein absorption. The results showed that cord mesenchymal stem cells and bone marrow mesenchymal stem cells had similar hepatocyte differentiation potential. However, compared with bone marrow mesenchymal stem cells, umbilical cord mesenchymal stem cells are easier to obtain and have low immunogenicity, so umbilical cord mesenchymal stem cells have more advantages as seed cells for tissue engineering. Polyhydroxyfatty acid ester (PHA) is a kind of macromolecule linear polyester produced by microorganisms in the environment of unbalanced nutrition. The research shows that PHA has good biocompatibility and biodegradability. Compared with other PHA materials, trimer PHBVHHx has better biocompatibility and more favorable for cell adhesion growth, so PHBVHHx has a good application prospect in liver tissue engineering. In this study we chose umbilical cord mesenchymal stem cells as seed cells of liver tissue engineering and trimer PHBVHHx as scaffold materials to study the effect of PHBVHHx scaffold on the differentiation of umbilical cord mesenchymal stem cells into hepatocytes in vitro. The results showed that PHBVHHx scaffold could support the growth of umbilical cord mesenchymal stem cells and that umbilical cord mesenchymal stem cells could differentiate into hepatoid cells induced by hepatocyte inducer on PHBVHHx scaffold. In a word, this study can provide some experimental information and data for liver tissue engineering.
【學(xué)位授予單位】：汕頭大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Human umbilical cord Wharton's Jelly-derived mesenchymal stem cells differentiation into nerve-like cells[J];Chinese Medical Journal;2005年23期

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本文編號：2416736