本文選題：人羊膜上皮細(xì)胞　切入點(diǎn)：腺泡細(xì)胞　出處：《遵義醫(yī)學(xué)院》2010年碩士論文

【摘要】： [目的]通過人羊膜上皮細(xì)胞(hAECs)體外誘導(dǎo)分化為腺泡細(xì)胞及人羊膜上皮細(xì)胞移植到受損傷唾液腺組織后的存活情況研究,為唾液腺功能障礙性疾病的細(xì)胞替代治療尋找理想的干細(xì)胞來源。 [方法]①采用機(jī)械法和酶消化法從人羊膜組織中分離hAECs,經(jīng)原代培養(yǎng)后用流式細(xì)胞儀(FCM)和免疫細(xì)胞化學(xué)方法進(jìn)行表型鑒定。②hAECs與SD大鼠唾液腺細(xì)胞用雙室裝置進(jìn)行共培養(yǎng)。③取誘導(dǎo)1周、2周后的hAECs,用免疫細(xì)胞化學(xué)、熒光實(shí)時(shí)定量RT-PCR等方法檢測α-淀粉酶及其mRNA的表達(dá)。④將BrdU標(biāo)記的hAECs懸液注入SD大鼠損傷下頜下腺組織內(nèi)。 [結(jié)果]①胰蛋白酶消化法能有效從人羊膜中分離hAECs,檢測結(jié)果顯示hAECs表達(dá)CD29和CK19陽性；CD44和α-淀粉酶弱陽性。②在誘導(dǎo)后2周內(nèi),hAECs中α-淀粉酶mRNA的表達(dá)量隨時(shí)間延長而增加。③免疫組化結(jié)果顯示,細(xì)胞移植后第4、7、14天組織切片均可見BrdU染色陽性細(xì)胞。[結(jié)論]hAECs具有向唾液腺樣細(xì)胞分化的能力,通過進(jìn)一步研究,可望成為唾液腺功能障礙性疾病細(xì)胞替代治療的干細(xì)胞來源。
[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 偽 -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 偽 -amylase weakly positive. 2 weeks after induction, the expression of 偽 -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.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

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