双室共培养中人羊膜上皮细胞诱导分化为腺泡细胞研究

发布时间：2018-04-05 01:37

本文选题：人羊膜上皮细胞　切入点：腺泡细胞　出处：《遵义医学院》2010年硕士论文

【摘要】： [目的]通过人羊膜上皮细胞(hAECs)体外诱导分化为腺泡细胞及人羊膜上皮细胞移植到受损伤唾液腺组织后的存活情况研究,为唾液腺功能障碍性疾病的细胞替代治疗寻找理想的干细胞来源。 [方法]①采用机械法和酶消化法从人羊膜组织中分离hAECs,经原代培养后用流式细胞仪(FCM)和免疫细胞化学方法进行表型鉴定。②hAECs与SD大鼠唾液腺细胞用双室装置进行共培养。③取诱导1周、2周后的hAECs,用免疫细胞化学、荧光实时定量RT-PCR等方法检测α-淀粉酶及其mRNA的表达。④将BrdU标记的hAECs悬液注入SD大鼠损伤下颌下腺组织内。 [结果]①胰蛋白酶消化法能有效从人羊膜中分离hAECs,检测结果显示hAECs表达CD29和CK19阳性；CD44和α-淀粉酶弱阳性。②在诱导后2周内,hAECs中α-淀粉酶mRNA的表达量随时间延长而增加。③免疫组化结果显示,细胞移植后第4、7、14天组织切片均可见BrdU染色阳性细胞。[结论]hAECs具有向唾液腺样细胞分化的能力,通过进一步研究,可望成为唾液腺功能障碍性疾病细胞替代治疗的干细胞来源。
[Abstract]:[objective] to study the survival of human amniotic epithelial cells induced to differentiate into acinar cells and human amniotic epithelial cells to the injured salivary glands in vitro.To find an ideal source of stem cells for cell replacement therapy for salivary gland dysfunction.[methods] 1 hAECs were isolated from human amniotic membrane tissue by mechanical method and enzyme digestion method. The phenotypes of hAECs and SD rat salivary gland cells were identified by flow cytometry (FCM) and immunocytochemistry after primary culture.After 1 week and 2 weeks of induction, hAECs were obtained by co-culture, and immunocytochemistry was used.The expression of 伪 -amylase and its mRNA was detected by real-time quantitative RT-PCR. 4. The BrdU labeled hAECs suspension was injected into the injured submandibular gland of SD rats.[results] 1 trypsin digestion method could effectively isolate hAECs from human amniotic membrane. The results showed that hAECs expressed CD29 and CK19 positive, CD44 and 伪 -amylase weakly positive. 2 weeks after induction, the expression of 伪 -amylase mRNA in hAECs was prolonged over time.Long and increased 3. 3 immunohistochemical results showed that,BrdU staining positive cells were found in tissue sections on the 4th day after transplantation.[conclusion] hAECs has the ability to differentiate into salivary gland like cells, and it is expected to be a stem cell source for the cell replacement therapy of salivary gland dysfunction by further study.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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