本文關(guān)鍵詞： 臍帶間充質(zhì)干細(xì)胞 豬角膜基質(zhì)后板層 紗布 凍存 血管內(nèi)皮細(xì)胞 組織工程　出處：《暨南大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：1.目的（1）提高體外分離hUC-MCs的成功率 （2）探討hUC-MCs作為組織工程角膜種子細(xì)胞的可能性 （3）改良凍存VECs的方法 （4）探討VECs作為組織工程角膜種子細(xì)胞的可能性 2.方法（1）用多片臍帶組織片懸浮法聯(lián)合Ⅳ型膠原酶消化法、單純多片臍帶組織片懸浮法及臍帶組織片懸浮法這三種方法體外分離hUC-MCs，并比較其成功率，鑒定細(xì)胞表面標(biāo)記并誘導(dǎo)其分化 （2）以豬角膜基質(zhì)后板層為載體培養(yǎng)hUC-MCs構(gòu)建組織工程角膜后板層并行動(dòng)物移植，觀察角膜透明情況，并行hUC-MCs表面特異內(nèi)皮標(biāo)志物NSE和ZO-1的檢測(cè) （3）采用改良及傳統(tǒng)方法凍存VECs，4周后復(fù)蘇分別比較其存活率、形態(tài)、生長(zhǎng)曲線，流式細(xì)胞術(shù)檢測(cè)其凋亡率 （4）以豬角膜基質(zhì)后板層為載體培養(yǎng)VECs構(gòu)建組織工程角膜后板層，掃描電鏡觀察其表面結(jié)構(gòu) 3.結(jié)果（1）多片懸浮臍帶片法聯(lián)合Ⅳ型膠原酶消化法及單純多片懸浮臍帶片法成功率高于懸浮臍帶片法 （2）種植hUC-MCs移植組角膜較非種植組透明，hUC-MCs表面特異內(nèi)皮標(biāo)志物NSE和ZO-1表達(dá)陽(yáng)性 （3）改良的凍存法VECs復(fù)蘇后形態(tài)上較傳統(tǒng)組好，存活率、生長(zhǎng)曲線及凋亡率與傳統(tǒng)組無(wú)明顯差異 (4）掃描電鏡可見(jiàn)hUC-MCs在豬角膜基質(zhì)后板層表面生長(zhǎng)良好，，連接成片 4.結(jié)論（1）本研究探討了hUC-MCs體外分離的幾種方法，提供了一種較為高效的培養(yǎng)方法 （2）hUC-MCs可向CECs方向轉(zhuǎn)化，為組織工程后板層的構(gòu)建提供了新的種子細(xì)胞來(lái)源 （3）改良的VECs凍存法能大幅度縮短凍存所需時(shí)間達(dá)到與傳統(tǒng)方法類似的效果 (4）VECs或能為組織工程后板層的構(gòu)建提供了新的種子細(xì)胞來(lái)源
[Abstract]:1. Objective 1) to improve the success rate of hUC-MCs isolation in vitro. To explore the possibility of hUC-MCs as a seed cell of tissue engineering cornea Improved method of VECs cryopreservation To explore the possibility of VECs as a seed cell of tissue engineering cornea 2. Methods hUC-MCs was isolated in vitro by multi-cord tissue suspension method combined with collagenase IV digestion method, simple multi-cord tissue suspension method and umbilical cord tissue suspension method in vitro. Compared with the success rate, the cell surface markers were identified and induced to differentiate. (2) using porcine corneal stromal posterior lamina (hUC-MCs) as the carrier to construct tissue engineered corneal posterior lamina and animal transplantation, the corneal transparency was observed. Detection of NSE and ZO-1 on hUC-MCs Surface specific Endothelial markers The survival rate, morphology, growth curve and apoptosis rate of VECs were measured by flow cytometry after 4 weeks resuscitation. VECs was used as the carrier to construct the tissue engineered posterior lamina of cornea, and the surface structure was observed by scanning electron microscope (SEM). 3. Results: the success rate of multislice suspension cord method combined with type 鈪

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