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  本文選題：成纖維細胞 + 細胞��； 參考：《山東大學》2006年碩士論文

【摘要】：目的：利用體外組織細胞培養(yǎng)及胚胎無瘢痕愈合的一般原理，在體外原代培養(yǎng)胚胎大鼠皮膚成纖維細胞，探索和建立大鼠胚胎皮膚成纖維細胞體外分離、培養(yǎng)及鑒定的技術方法。觀察大鼠胚胎皮膚成纖維細胞在正常皮膚以及深二度燒傷創(chuàng)面注射移植后的存活情況。 方法：分別用組織塊法和酶消化分離法原代培養(yǎng)處于無瘢痕愈合期的Wistar大鼠胚胎皮膚成纖維細胞，對兩種方法獲得細胞的情況進行比較。用胰蛋白酶+EDTA消化加反復貼壁法分離純化成纖維細胞。倒置相差顯微鏡下觀察原代培養(yǎng)及傳代過程中成纖維細胞的形態(tài)學特征。制備細胞爬片，對培養(yǎng)的成纖維細胞用SP方法作免疫細胞化學染色(immunocytochemistry，ICC)，第一抗體分別為波形蛋白(vimentin)、結蛋白(desmin)、角蛋白(pan cytokeratin)。選取生長狀態(tài)良好的細胞，制作標本，用透射電子顯微鏡(transmission electron microscope，TEM)觀察胚胎成纖維細胞的超微結構。 用組織塊法和酶消化法對成年Wistar大鼠皮膚成纖維細胞進行培養(yǎng)，并對兩類細胞生長以及功能活動情況進行簡單的比較研究。細胞計數(shù)法繪制兩種細胞的生長曲線。流式細胞術(flow cytometry，F(xiàn)CM)測定比較處于對數(shù)生長期的兩種細胞的細胞周期情況，細胞各周期的百分比測定結果以(?)±S表示，作組間t檢驗進行統(tǒng)計學分析，，P＜0.05即認為有統(tǒng)計學意義。 選擇成年健康Wistar大鼠12只，每只體重約250g左右，隨機分為兩組，A組為正常皮膚移植組，B組為燒傷創(chuàng)面移植組。A組每只背
[Abstract]:Objective : To explore and establish a technique for the in vitro isolation , culture and identification of rat embryonic skin fibroblasts by using the general principles of in vitro tissue culture and embryo - free healing , and to observe the survival of rat embryonic skin fibroblasts after injection transplantation in normal skin and deep second degree burn wounds .










Methods : The embryonic skin fibroblasts were cultured in Wistar rats without scar healing stage by tissue block method and enzyme digestion separation method . The cells were isolated and purified by trypsin + EDTA digestion and repeated adherent method . The cells were cultured and passaged by reversed phase contrast microscope . The cells were stained by SP method for immunohistochemical staining ( ICC ) . The first antibody was vimentin , vimentin , pan cytokeratin , respectively . The ultrastructure of embryonic fibroblasts was observed by transmission electron microscope ( TEM ) .










The growth curve of two kinds of cells was studied by using tissue block method and enzymatic digestion method . The growth curve of two kinds of cells was studied by cell counting method . Flow cytometry ( FCM ) was used to measure the cell cycle of two kinds of cells .










12 adult healthy Wistar rats were randomly divided into two groups : group A , normal skin transplantation group , group B as normal skin graft group and group B as burn wound transplantation group .

【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R329.2

【引證文獻】

相關期刊論文 前1條

1 石慧娟;黃大元;;成纖維細胞及其相關因素在無瘢痕愈合中的研究進展[J];四川解剖學雜志;2011年01期

本文編號：1883640