發(fā)布時(shí)間：2018-06-01 20:39

  本文選題：胰島素 + 小鼠��； 參考：《昆明醫(yī)學(xué)院》2008年碩士論文

【摘要】： 【目的】研究胰島素(Insulin)對(duì)ICR小鼠早期胚胎體外發(fā)育的影響,以探討胰島素在小鼠早期胚胎體外發(fā)育中的作用,為進(jìn)一步優(yōu)化胚胎體外培養(yǎng)體系提供理論和實(shí)驗(yàn)依據(jù)。 【方法】取ICR小鼠胚胎隨機(jī)分為實(shí)驗(yàn)組與對(duì)照組在體外連續(xù)培養(yǎng)(所用基礎(chǔ)培養(yǎng)基為mKSOM),觀察囊胚率、孵化率和囊胚細(xì)胞數(shù)。實(shí)驗(yàn)一:觀察不同濃度胰島素對(duì)小鼠1-細(xì)胞胚胎體外發(fā)育的影響。實(shí)驗(yàn)共分六組:1組為對(duì)照組,培養(yǎng)基中不添加胰島素。2、3、4、5、6組為實(shí)驗(yàn)組,分別在培養(yǎng)基中添加0.005、0.05、0.5、5和10 ug/ml濃度的胰島素。實(shí)驗(yàn)二:觀察胰島素對(duì)小鼠早期胚胎體外發(fā)育各階段的影響。實(shí)驗(yàn)共分六組:1組為對(duì)照組,培養(yǎng)全程均不含胰島素。其余為實(shí)驗(yàn)組,2、3、4、5組分別在1-細(xì)胞、2-細(xì)胞、4-細(xì)胞及桑椹胚階段移入含0.05ug/ml胰島素的培養(yǎng)基,24小時(shí)后又移到無(wú)胰島素的培養(yǎng)基中;6組為培養(yǎng)全程均含0.05 ug/ml胰島素。實(shí)驗(yàn)三:觀察不同時(shí)期(2-細(xì)胞期、4-細(xì)胞期)小鼠胚胎體外培養(yǎng)所需胰島素的濃度。每個(gè)實(shí)驗(yàn)均分六組:1組為對(duì)照組,培養(yǎng)基中不添加胰島素。2、3、4、5、6組為實(shí)驗(yàn)組,分別在培養(yǎng)基中添加0.005、0.05、0.5、5和10 ug/ml濃度的胰島素。 【結(jié)果】實(shí)驗(yàn)一:1)、實(shí)驗(yàn)組與對(duì)照組比較,囊胚率、孵化率及囊胚細(xì)胞數(shù)顯著提高(P＜0.05);2)、五個(gè)實(shí)驗(yàn)組間比較,囊胚率、孵化率沒有差別(P＞0.05);除組2和組3(即胰島素濃度為0.005 ug/ml、0.05 ug/ml組)與其余實(shí)驗(yàn)組比較,囊胚細(xì)胞數(shù)提高(P＜0.05)外,其余各實(shí)驗(yàn)組間囊胚細(xì)胞數(shù)沒有差別(P＞0.05);組2與組3間囊胚細(xì)胞數(shù)沒有差別(P＞0.05)。 實(shí)驗(yàn)二:1)、2-細(xì)胞、4-細(xì)胞階段移入含胰島素的培養(yǎng)基及培養(yǎng)基中一直含胰島素組與對(duì)照組及其余實(shí)驗(yàn)組比較,囊胚率及囊胚細(xì)胞數(shù)顯著提高(P＜0.05);2)、4-細(xì)胞階段移入含胰島素的培養(yǎng)基與對(duì)照組及其余實(shí)驗(yàn)組比較,孵化率顯著提高(P＜0.05);3)、1-細(xì)胞、桑椹胚階段移入含胰島素的培養(yǎng)基與對(duì)照組及其余實(shí)驗(yàn)組比較,囊胚率、孵化率及囊胚細(xì)胞數(shù)沒有差別(P＞0.05)。 實(shí)驗(yàn)三:共分兩部分:第一部分(2-細(xì)胞期添加胰島素):1)、各濃度實(shí)驗(yàn)組與未加胰島素對(duì)照組比較,囊胚率、孵化率及囊胚細(xì)胞數(shù)顯著提高(p＜0.05);2)、五個(gè)實(shí)驗(yàn)組間比較,囊胚率、孵化率沒有差別(P＞0.05);除組2和組3與其余實(shí)驗(yàn)組比較,囊胚細(xì)胞數(shù)顯著提高(P＜0.05)外,其余各實(shí)驗(yàn)組間囊胚細(xì)胞數(shù)沒有差別(P＞0.05);組2與組3比較,囊胚細(xì)胞數(shù)沒有差別(P＞0.05)。第二部分(4-細(xì)胞期添加胰島素):1)、各濃度實(shí)驗(yàn)組與未加胰島素對(duì)照組比較,囊胚率、孵化率及囊胚細(xì)胞數(shù)顯著提高(p＜0.05);2)、五個(gè)實(shí)驗(yàn)組間比較,囊胚率、孵化率沒有差別(P＞0.05);除組3與其余實(shí)驗(yàn)組比較,囊胚細(xì)胞數(shù)顯著提高(P＜0.05)外,其余各實(shí)驗(yàn)組間囊胚細(xì)胞數(shù)比較沒有差別(P＞0.05)。 【結(jié)論】1、體外培養(yǎng)時(shí),在培養(yǎng)基中添加胰島素對(duì)ICR小鼠早期胚胎發(fā)育有促進(jìn)作用。2、2-細(xì)胞及4-細(xì)胞階段添加胰島素,對(duì)早期胚胎發(fā)育有促進(jìn)作用。3、胚胎發(fā)育的不同階段對(duì)胰島素的需求有所不同,0.05 ug/ml為2-細(xì)胞、4-細(xì)胞階段體外培養(yǎng)時(shí)的最佳濃度。4、胰島素濃度在0.005～10 ug/ml范圍內(nèi),對(duì)體外培養(yǎng)的小鼠胚胎無(wú)毒性作用。
[Abstract]:[Objective] to study the effect of insulin (Insulin) on the development of early embryos of ICR mice in vitro, in order to explore the role of insulin in the development of early mouse embryos in vitro, and to provide theoretical and experimental basis for further optimizing the culture system of embryos in vitro.
[Methods] the ICR mouse embryos were randomly divided into the experimental group and the control group in vitro (the base medium was mKSOM), and the blastocyst rate, the hatchability and the number of blastocysts were observed. Experiment 1: the effect of insulin on the development of mouse 1- cell embryos in vitro was observed. The results were divided into six groups: the 1 groups were the control group, and the culture medium did not add. The insulin.2,3,4,5,6 group was in the experimental group, adding 0.005,0.05,0.5,5 and 10 ug/ml concentration of insulin in the medium. Experiment two: the effect of insulin on the development of the early embryos in mice was observed. The experiment was divided into six groups: the 1 groups were in the control group and the whole course was no isisin. The rest was the experimental group, and the 2,3,4,5 group was in 1- cells, respectively. 2- cells, 4- cells and morula stage were moved into the medium containing 0.05ug/ml insulin and moved to the non insulin medium after 24 hours. The 6 groups were 0.05 ug/ml insulin in the whole course of culture. Experiment three: the concentration of insulin needed in the culture of mice embryos in different periods (2- cell stage, 4- cell stage). Each experiment was divided into six groups: 1 groups In the control group, the.2,3,4,5,6 group without insulin was added to the medium. 0.005,0.05,0.5,5 and 10 ug/ml of insulin were added to the medium respectively.
[results] Experiment 1: 1), compared with the control group, the rate of blastocyst, hatchability and the number of blastocysts were significantly increased (P < 0.05), 2). There was no difference between the blastocyst rate and the hatching rate between the five experimental groups (P > 0.05), and the number of blastocysts in group 2 and group 3 (i.e., insulin concentration 0.005 ug/ml, 0.05 ug/ml group) increased (P < 0). 5) there was no difference in the number of blastocyst cells between the other experimental groups (P > 0.05); there was no difference in the number of blastocyst cells between group 2 and group 3 (P > 0.05).
Experiments two: 1), 2- cells, 4- cell stage moved into the medium containing insulin and the medium of insulin and the control group and the other experimental group, the blastocyst rate and the number of blastocyst cells increased significantly (P < 0.05), 2). The incubation rate of the 4- cell stage moved into the insulin containing medium and the control group and the rest of the experimental group, the hatching rate was significantly increased (P 0.05); (3), 1- cells, there was no difference in blastocyst rate, hatchability and number of blastocysts (P > 0.05) compared with the control group and the other experimental groups when the morula stage was moved into the culture medium containing insulin.
Experiment three: the first part was divided into two parts: the first part (2- cell phase adding insulin): 1), the blastocyst rate, the hatchability and the number of blastocysts were significantly increased (P < 0.05) in the experimental group and the non insulin control group (2). The rate of blastocyst and the hatching rate were no difference between the five experimental groups (P > 0.05); except for group 2 and group 3, the blastocyst was compared with the other experimental groups. There was no difference in the number of blastocysts (P > 0.05) among the other experimental groups (P < 0.05). Compared with group 3, the number of blastocyst cells did not differ (P > 0.05). The second part (4- cell phase added insulin) was 1). The blastocyst rate, hatchability and blastocyst cell number of the experimental group were significantly higher than those in the non islet control group (P < 0.05); 2), compared with the five experimental groups, there was no difference in blastocyst rate and hatchability (P > 0.05). Except for group 3, the number of blastocysts in the rest of the experimental groups was significantly increased (P < 0.05), and there was no difference in the number of blastocyst cells between the other experimental groups (P > 0.05).
[Conclusion] 1, when cultured in vitro, insulin added to the early embryo development of ICR mice was added to the.2,2- cell and 4- cell stage in the culture medium. The early embryo development was promoted by.3. The different stages of embryo development were different to the insulin demand, 0.05 ug/ml was 2- cell, and the 4- cell stage was cultured in vitro. The optimum concentration of.4 and the concentration of insulin in the range of 0.005 to 10 ug/ml had no toxic effect on mouse embryos in vitro.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R321-33

【引證文獻(xiàn)】

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本文編號(hào)：1965550