【摘要】： 目的：通過觀察原代昆明小鼠胰腺導(dǎo)管上皮樣細(xì)胞在動態(tài)培養(yǎng)條件下的分化狀態(tài)以及該分化后的細(xì)胞分泌胰島素的功能，探討正常昆明小鼠胰腺導(dǎo)管上皮樣細(xì)胞向胰島素分泌細(xì)胞分化的誘導(dǎo)條件、誘導(dǎo)后的胰島素分泌細(xì)胞分泌胰島素的功能以及分泌的胰島素的降糖活性，為1型糖尿病的細(xì)胞替代治療提供實(shí)驗(yàn)依據(jù)。 方法：取正常成年昆明小鼠胰腺導(dǎo)管上皮進(jìn)行原代培養(yǎng)，利用細(xì)胞貼壁時間的差異在培養(yǎng)24 h、48 h、72 h連續(xù)換液除去腺體細(xì)胞，在含2％胎牛血清的培養(yǎng)條件下除去成纖維細(xì)胞，，使可以在無血清培養(yǎng)基中生長的胰腺導(dǎo)管上皮樣細(xì)胞形成優(yōu)勢生長。在此條件下培養(yǎng)、傳代并鑒定。取第三代細(xì)胞以無血清誘導(dǎo)培養(yǎng)基促使其分化。取誘導(dǎo)后3 d、5 d、7 d、14 d的細(xì)胞進(jìn)行免疫細(xì)胞化學(xué)染色鑒定。ELISA檢測誘導(dǎo)生成的胰島素分泌細(xì)胞在高糖刺激下分泌胰島素的功能。用高糖培養(yǎng)基刺激誘導(dǎo)生成的胰島素分泌細(xì)胞后離心取上清液注射到正常昆明小鼠腹部皮下，檢測所分泌的胰島素的降糖活性。 結(jié)果：經(jīng)過無血清誘導(dǎo)培養(yǎng)基誘導(dǎo)7 d后，胰腺導(dǎo)管上皮樣細(xì)胞開始向胰島素分泌細(xì)胞分化，胰島素抗體染色陽性。14 d胰島素抗體染色陽性細(xì)胞明顯增多。該細(xì)胞在高糖刺激下可以分泌胰島素。所分泌的胰島素可以使正常昆明小鼠的血糖下降。 結(jié)論：昆明小鼠胰腺導(dǎo)管上皮樣細(xì)胞經(jīng)過無血清培養(yǎng)基誘導(dǎo)可以向胰島素分泌細(xì)胞方向分化。并且該胰島素分泌細(xì)胞在高糖刺激下具有分泌胰島素的功能。所分泌的胰島素具有降糖活性。
[Abstract]:Aim: to observe the differentiation of primary pancreatic ductal epithelioid cells in Kunming mice under dynamic culture and the function of insulin secretion by the differentiated cells. To investigate the inducing conditions of differentiation of pancreatic ductal epithelioid cells into insulin secreting cells in normal Kunming mice, the function of insulin secretion cells and the hypoglycemic activity of insulin secreted by induced insulin secretion cells. To provide experimental evidence for cell replacement therapy of type 1 diabetes. Methods: the pancreatic ductal epithelium of normal adult Kunming mice was cultured in primary culture. The glandular cells were removed from the pancreatic ductal epithelium at 24 h, 48 h and 72 h by cell adhesion time. The fibroblasts were removed under the condition of 2% fetal bovine serum, and the pancreatic ductal epithelioid cells which could grow in serum-free medium formed dominant growth. In this condition, culture, passage and identification. The third generation cells were induced to differentiate by serum-free medium. The immunocytochemical staining was performed on the cells at 3 days, 5 days, 7 days and 14 days after induction. ELISA was used to detect the function of insulin secreting cells induced by high glucose stimulation. The insulin secreting cells induced by high glucose medium were then centrifuged and injected into the abdominal subcutaneous of normal Kunming mice to detect the hypoglycemic activity of the insulin secreted. Results: after induction on serum-free medium for 7 days, pancreatic ductal epithelioid cells began to differentiate into insulin-secreting cells, and insulin antibody staining positive cells increased significantly at 14 days. The cells secrete insulin under high glucose stimulation. The insulin secreted can reduce the blood sugar of normal Kunming mice. Conclusion: pancreatic ductal epithelioid cells of Kunming mice can differentiate into insulin secreting cells induced by serum-free medium. Furthermore, the insulin secreting cells have the function of secreting insulin under high glucose stimulation. The insulin secreted has hypoglycemic activity.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 史愷;徐立;鄧儀昊;陳道邦;;昆明小鼠胰腺導(dǎo)管上皮樣細(xì)胞向胰島樣細(xì)胞的誘導(dǎo)分化[J];四川解剖學(xué)雜志;2007年01期

2 馮仁青;糖尿病的細(xì)胞治療[J];中國生物工程雜志;2003年07期

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