妊娠對脂肪干細胞增殖活性的影響
發(fā)布時間：2018-09-02 09:13
【摘要】：[目的]本實驗采用組織工程學原理,提取不同妊娠時期的ADSCs進行培養(yǎng)、鑒定,然后對其增殖活性進行對比分析,同時檢測雌孕激素對ADSCs增殖活性的影響。如實驗結(jié)果符合預(yù)期,提取妊娠期干細胞作為種子細胞進行培養(yǎng)將可能取得良好效果,同時也是對其生物學特性的有益探索。 [方法](1)完成對ADSCs的分離培養(yǎng),并適時對細胞進行傳代,采用光鏡觀察細胞形態(tài)和狀態(tài)；(2)通過對ADSCs表面抗原的檢測及成骨誘導實驗完成對ADSCs的鑒定；(3)選取不同分組的第3代ADSCs,通過MTT法和碘化丙錠(PI)染色分別檢測各組吸光度值和細胞周期,繪制細胞生長曲線,并對檢測結(jié)果進行統(tǒng)計學分析,對比不同組ADSCs增殖活性的差異。 [結(jié)果](1)各組細胞生長良好,呈梭形旋渦狀增殖,組織分離24h后妊娠組貼壁的梭形細胞數(shù)量較未孕組多；(2)細胞表面抗原檢測結(jié)果顯示所培養(yǎng)細胞CD90+CD44+CD34-,成骨誘導結(jié)果顯示有紅色鈣結(jié)節(jié)生成；(3)MTT實驗結(jié)果顯示：未孕ADSCs+P組與未孕ADSCs組,孕中期ADSCs組、臨產(chǎn)ADSCs組、未孕ADSCs+E2組組間差異無統(tǒng)計學意義(P0.05)；孕中期ADSCs組、臨產(chǎn)ADSCs組、未孕ADSCs+E2組與未孕ADSCs組相比差異有統(tǒng)計學意義(P0.05)；(4)PI染色檢測各組細胞周期顯示：孕中期ADSCs組,臨產(chǎn)ADSCs組和普通ADSCs+E2組與普通ADSCs組相比,處于非增殖周期的Go/G1期細胞所占比例呈下降趨勢,差異有統(tǒng)計學意義(p0.05)；未孕ADSCs+P組與未孕ADSCs組,孕中期ADSCs組與臨產(chǎn)ADSCs組相比處于Go/G,期的細胞所占比例無明顯增減,差異無統(tǒng)計學意義(p0.05)。 [結(jié)論]所培養(yǎng)的細胞為ADSCs,且在體外培養(yǎng)環(huán)境下生長良好；妊娠大鼠的ADSCs增殖活性較未妊娠者高,妊娠中期與臨產(chǎn)期ADSCs增殖活性無明顯差異；雌二醇能促進ADSCs的增殖,孕酮則未表現(xiàn)出促進ADSCs增殖的能力。
[Abstract]:[objective] to investigate the effects of estrogen and progesterone on the proliferation of ADSCs in different pregnancy stages by using tissue engineering principle, and then compared and analyzed its proliferative activity. If the experimental results are in line with the expectation, it is possible to obtain good results by extracting pregnancy stem cells as seed cells, and it is also a useful exploration of their biological characteristics. [methods] (1) the ADSCs was isolated and cultured, and the cells were subcultured at the right time. The morphology and state of the cells were observed by light microscope. (2) the ADSCs was identified by the detection of ADSCs surface antigen and the osteogenesis induction experiment. (3) the third generation ADSCs, of different groups were selected to detect the absorbance and cell cycle of each group by MTT method and (PI) staining respectively, and to draw the cell growth curve. The results were analyzed statistically to compare the proliferative activity of different groups of ADSCs. [results] (1) the cells in each group grew well and proliferated in fusiform swirl shape. After 24 hours of tissue separation, the number of fusiform cells adhered to the wall in pregnancy group was more than that in non-pregnant group. (2) the results of cell surface antigen test showed that the CD90 CD44 CD34-, osteogenesis of cultured cells showed red calcium nodule formation, (3) MTT test showed that: ADSCs P group, ADSCs group. There was no significant difference between non-pregnant ADSCs E2 group (P0.05), second trimester ADSCs group, parturient ADSCs group, non-pregnant ADSCs E2 group and non-pregnant ADSCs group had significant difference (P0.05); (4) PI staining showed that the second trimester ADSCs group, the non-pregnant ADSCs E2 group and the non-pregnant ADSCs group had significant difference (P0.05); (4). The percentage of Go/G1 cells in non-proliferative cycle in ADSCs group and ADSCs E2 group decreased significantly (p0.05), and ADSCs P group and non-pregnant ADSCs group showed significant difference (p0.05), the percentage of Go/G1 phase cells in non-proliferative cycle group was significantly lower than that in normal ADSCs group (p0.05), while that in non-pregnant ADSCs P group and non-pregnant ADSCs group was significantly lower (p0.05). The percentage of cells in Go/G, phase in ADSCs group was not significantly increased compared with that in parturient ADSCs group (p0.05). [conclusion] the cultured cells were ADSCs, and grew well in vitro, the proliferative activity of ADSCs in pregnant rats was higher than that in unpregnant rats, and the proliferative activity of ADSCs in the second trimester of pregnancy was not significantly different from that in parturient stage, estradiol could promote the proliferation of ADSCs. Progesterone did not show the ability to promote the proliferation of ADSCs.
【學位授予單位】：昆明醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R329
本文編號：2218910
[Abstract]:[objective] to investigate the effects of estrogen and progesterone on the proliferation of ADSCs in different pregnancy stages by using tissue engineering principle, and then compared and analyzed its proliferative activity. If the experimental results are in line with the expectation, it is possible to obtain good results by extracting pregnancy stem cells as seed cells, and it is also a useful exploration of their biological characteristics. [methods] (1) the ADSCs was isolated and cultured, and the cells were subcultured at the right time. The morphology and state of the cells were observed by light microscope. (2) the ADSCs was identified by the detection of ADSCs surface antigen and the osteogenesis induction experiment. (3) the third generation ADSCs, of different groups were selected to detect the absorbance and cell cycle of each group by MTT method and (PI) staining respectively, and to draw the cell growth curve. The results were analyzed statistically to compare the proliferative activity of different groups of ADSCs. [results] (1) the cells in each group grew well and proliferated in fusiform swirl shape. After 24 hours of tissue separation, the number of fusiform cells adhered to the wall in pregnancy group was more than that in non-pregnant group. (2) the results of cell surface antigen test showed that the CD90 CD44 CD34-, osteogenesis of cultured cells showed red calcium nodule formation, (3) MTT test showed that: ADSCs P group, ADSCs group. There was no significant difference between non-pregnant ADSCs E2 group (P0.05), second trimester ADSCs group, parturient ADSCs group, non-pregnant ADSCs E2 group and non-pregnant ADSCs group had significant difference (P0.05); (4) PI staining showed that the second trimester ADSCs group, the non-pregnant ADSCs E2 group and the non-pregnant ADSCs group had significant difference (P0.05); (4). The percentage of Go/G1 cells in non-proliferative cycle in ADSCs group and ADSCs E2 group decreased significantly (p0.05), and ADSCs P group and non-pregnant ADSCs group showed significant difference (p0.05), the percentage of Go/G1 phase cells in non-proliferative cycle group was significantly lower than that in normal ADSCs group (p0.05), while that in non-pregnant ADSCs P group and non-pregnant ADSCs group was significantly lower (p0.05). The percentage of cells in Go/G, phase in ADSCs group was not significantly increased compared with that in parturient ADSCs group (p0.05). [conclusion] the cultured cells were ADSCs, and grew well in vitro, the proliferative activity of ADSCs in pregnant rats was higher than that in unpregnant rats, and the proliferative activity of ADSCs in the second trimester of pregnancy was not significantly different from that in parturient stage, estradiol could promote the proliferation of ADSCs. Progesterone did not show the ability to promote the proliferation of ADSCs.
【學位授予單位】：昆明醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R329
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