胰岛素对ICR小鼠早期胚胎体外发育的影响

发布时间：2018-06-01 20:39

本文选题：胰岛素 + 小鼠　；参考：《昆明医学院》2008年硕士论文

【摘要】： 【目的】研究胰岛素(Insulin)对ICR小鼠早期胚胎体外发育的影响,以探讨胰岛素在小鼠早期胚胎体外发育中的作用,为进一步优化胚胎体外培养体系提供理论和实验依据。【方法】取ICR小鼠胚胎随机分为实验组与对照组在体外连续培养(所用基础培养基为mKSOM),观察囊胚率、孵化率和囊胚细胞数。实验一:观察不同浓度胰岛素对小鼠1-细胞胚胎体外发育的影响。实验共分六组:1组为对照组,培养基中不添加胰岛素。2、3、4、5、6组为实验组,分别在培养基中添加0.005、0.05、0.5、5和10 ug/ml浓度的胰岛素。实验二:观察胰岛素对小鼠早期胚胎体外发育各阶段的影响。实验共分六组:1组为对照组,培养全程均不含胰岛素。其余为实验组,2、3、4、5组分别在1-细胞、2-细胞、4-细胞及桑椹胚阶段移入含0.05ug/ml胰岛素的培养基,24小时后又移到无胰岛素的培养基中;6组为培养全程均含0.05 ug/ml胰岛素。实验三:观察不同时期(2-细胞期、4-细胞期)小鼠胚胎体外培养所需胰岛素的浓度。每个实验均分六组:1组为对照组,培养基中不添加胰岛素。2、3、4、5、6组为实验组,分别在培养基中添加0.005、0.05、0.5、5和10 ug/ml浓度的胰岛素。【结果】实验一:1)、实验组与对照组比较,囊胚率、孵化率及囊胚细胞数显著提高(P＜0.05);2)、五个实验组间比较,囊胚率、孵化率没有差别(P＞0.05);除组2和组3(即胰岛素浓度为0.005 ug/ml、0.05 ug/ml组)与其余实验组比较,囊胚细胞数提高(P＜0.05)外,其余各实验组间囊胚细胞数没有差别(P＞0.05);组2与组3间囊胚细胞数没有差别(P＞0.05)。实验二:1)、2-细胞、4-细胞阶段移入含胰岛素的培养基及培养基中一直含胰岛素组与对照组及其余实验组比较,囊胚率及囊胚细胞数显著提高(P＜0.05);2)、4-细胞阶段移入含胰岛素的培养基与对照组及其余实验组比较,孵化率显著提高(P＜0.05);3)、1-细胞、桑椹胚阶段移入含胰岛素的培养基与对照组及其余实验组比较,囊胚率、孵化率及囊胚细胞数没有差别(P＞0.05)。实验三:共分两部分:第一部分(2-细胞期添加胰岛素):1)、各浓度实验组与未加胰岛素对照组比较,囊胚率、孵化率及囊胚细胞数显著提高(p＜0.05);2)、五个实验组间比较,囊胚率、孵化率没有差别(P＞0.05);除组2和组3与其余实验组比较,囊胚细胞数显著提高(P＜0.05)外,其余各实验组间囊胚细胞数没有差别(P＞0.05);组2与组3比较,囊胚细胞数没有差别(P＞0.05)。第二部分(4-细胞期添加胰岛素):1)、各浓度实验组与未加胰岛素对照组比较,囊胚率、孵化率及囊胚细胞数显著提高(p＜0.05);2)、五个实验组间比较,囊胚率、孵化率没有差别(P＞0.05);除组3与其余实验组比较,囊胚细胞数显著提高(P＜0.05)外,其余各实验组间囊胚细胞数比较没有差别(P＞0.05)。【结论】1、体外培养时,在培养基中添加胰岛素对ICR小鼠早期胚胎发育有促进作用。2、2-细胞及4-细胞阶段添加胰岛素,对早期胚胎发育有促进作用。3、胚胎发育的不同阶段对胰岛素的需求有所不同,0.05 ug/ml为2-细胞、4-细胞阶段体外培养时的最佳浓度。4、胰岛素浓度在0.005～10 ug/ml范围内,对体外培养的小鼠胚胎无毒性作用。
[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.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R321-33

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