人脐血造血干细胞小鼠移植模型的建立
发布时间：2019-01-22 10:10
【摘要】： 造血干细胞移植是治疗血液及免疫系统等疾病的主要手段,通过造血干细胞移植也可以为多种疾病提供动物模型,尤其是人造血干细胞移植小鼠建立的人源化血液系统小鼠模型为研究某些无法感染小鼠的人类疾病病毒提供有效途径。本研究从脐血中分离出人CD34阳性造血干细胞,分别通过骨髓移植、新生肝移植、宫内移植方法建立人源化造血系统小鼠模型。在小鼠移植后1个月至3个月检测人血细胞表面抗原CD45。新生肝移植后同时检测了髓系细胞表面抗原CD33、T淋巴细胞表面抗原CD3以及B淋巴细胞表面抗原CD19。通过人血细胞表面抗原的表达检测出小鼠体内人血液系统的重建情况。 1、利用骨髓移植方法共移植12只NOD/SCID小鼠,阴性对照组4只,试验组8只。试验组小鼠均取得了人血液细胞的重建,1个月后检测外周血人CD45得到46.88%的重建率,3个月后骨髓中人血细胞重建率达到96.24%。试验结果证明通过骨髓移植人造血干细胞可以成功促进人造血细胞在小鼠体内的重建。 2、通过新生肝移植方法共移植两窝16只NOD/SCID仔鼠,阴性对照组4只,试验组12只。阴性对照小鼠均在2周内死亡,未检测到人血细胞重建;试验组小鼠均检测到人血液细胞重建,有三只小鼠存活两个月以上。1个月后检测外周血人CD45得到26.8%的重建率,2个月后外周血重建率达到60.94%,骨髓重建率高达86.49%。2个月后脾脏内的人T、B、髓系细胞分别占4.29%、47.17%和5.4%;骨髓内的T、B、髓系细胞分别占0.25%、54.27%和23.5%;外周血内各系血细胞占的比例较低,T、B、髓系细胞分别占0.7%、26.42%和2.69%。试验结果证明了通过新生肝移植方法能够得到高效、全面的造血重建。 3、在E8.5d宫内卵黄囊移植试验中,共获得健康出生仔鼠22只,除一只惺笾亟缡蚀锏?.34%,其他检测结果基本处于1%以下。通过E12.5d宫内腹腔移植得到健康出生小鼠8只,其中6只检测到人CD45阳性表达,但重建率总体低于卵黄囊移植得到的重建率。本研究同时进行了NOD/SCID胎鼠卵黄囊移植,获得健康出生的仔鼠11只,但重建比例亦低于1%。试验结果说明卵黄囊移植优于腹腔移植,但通过宫内移植得到的重建率低于其他移植方式。另外,本研究未发现免疫缺陷小鼠宫内移植对于造血重建的优越性。
[Abstract]:Hematopoietic stem cell transplantation (HSCT) is the main method for the treatment of diseases such as blood and immune system, and it can also provide animal models for many diseases. Especially the artificial blood stem cell transplantation mouse model of humanized blood system provides an effective way to study some human disease virus which can not be infected in mice. In this study, human CD34 positive hematopoietic stem cells were isolated from umbilical cord blood, and human hematopoietic system models were established by bone marrow transplantation, new liver transplantation and intrauterine transplantation respectively. Detection of human blood cell surface antigen CD45. from 1 to 3 months after transplantation in mice CD33,T lymphocyte surface antigen (CD3) and B lymphocyte surface antigen (CD19.) of myeloid cell surface antigen (CD33,T) were detected after liver transplantation. The reconstitution of human blood system in mice was detected by the expression of human blood cell surface antigen. 1. A total of 12 NOD/SCID mice were transplanted by bone marrow transplantation, 4 in negative control group and 8 in experimental group. Human blood cells were reconstructed in all mice in the experimental group. The reconstruction rate of human CD45 in peripheral blood was 46.88% after one month and 96.2424% in bone marrow 3 months later. The results show that bone marrow transplantation of artificial blood stem cells can successfully promote the reconstruction of human hematopoietic cells in mice. 2. Two litters of 16 NOD/SCID mice were transplanted by the method of neonate liver transplantation, 4 rats in negative control group and 12 rats in test group. All the negative control mice died within 2 weeks and no human blood cells were detected. Three mice survived for more than two months. After one month, the reconstruction rate of human CD45 in peripheral blood was 26.8%, and the reconstruction rate of peripheral blood reached 60.94% 2 months later. The rate of bone marrow reconstruction was as high as 86.49.2 months later, and the percentage of medullary cells in spleen was 47.17% and 5.4%, respectively. In bone marrow, medullary cells accounted for 54.27% and 23.5%, respectively, and the percentage of blood cells in peripheral blood was lower, and the percentage of TnB and myeloid cells was 0.726.42% and 2.69%, respectively. The results show that the new liver transplantation method can achieve high efficiency and comprehensive hematopoietic reconstitution. 3. In E8.5 days intrauterine yolk sac transplantation test, 22 healthy newborn rats were obtained. Except for one mouse, the other results were basically below 1%. Eight healthy born mice were obtained by intrauterine transplantation of E12.5 days, 6 of which detected positive expression of human CD45, but the reconstruction rate was lower than that of yolk sac transplantation. In this study, NOD/SCID fetal yolk sac transplantation was performed and 11 healthy offspring were obtained, but the reconstruction rate was less than 1. The results showed that yolk sac transplantation was superior to abdominal transplantation, but the reconstruction rate of intrauterine transplantation was lower than that of other transplantation methods. In addition, the advantage of intrauterine transplantation for hematopoietic reconstitution in immunodeficient mice was not found in this study.
【学位授予单位】：内蒙古大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R-332
本文编号：2413122
[Abstract]:Hematopoietic stem cell transplantation (HSCT) is the main method for the treatment of diseases such as blood and immune system, and it can also provide animal models for many diseases. Especially the artificial blood stem cell transplantation mouse model of humanized blood system provides an effective way to study some human disease virus which can not be infected in mice. In this study, human CD34 positive hematopoietic stem cells were isolated from umbilical cord blood, and human hematopoietic system models were established by bone marrow transplantation, new liver transplantation and intrauterine transplantation respectively. Detection of human blood cell surface antigen CD45. from 1 to 3 months after transplantation in mice CD33,T lymphocyte surface antigen (CD3) and B lymphocyte surface antigen (CD19.) of myeloid cell surface antigen (CD33,T) were detected after liver transplantation. The reconstitution of human blood system in mice was detected by the expression of human blood cell surface antigen. 1. A total of 12 NOD/SCID mice were transplanted by bone marrow transplantation, 4 in negative control group and 8 in experimental group. Human blood cells were reconstructed in all mice in the experimental group. The reconstruction rate of human CD45 in peripheral blood was 46.88% after one month and 96.2424% in bone marrow 3 months later. The results show that bone marrow transplantation of artificial blood stem cells can successfully promote the reconstruction of human hematopoietic cells in mice. 2. Two litters of 16 NOD/SCID mice were transplanted by the method of neonate liver transplantation, 4 rats in negative control group and 12 rats in test group. All the negative control mice died within 2 weeks and no human blood cells were detected. Three mice survived for more than two months. After one month, the reconstruction rate of human CD45 in peripheral blood was 26.8%, and the reconstruction rate of peripheral blood reached 60.94% 2 months later. The rate of bone marrow reconstruction was as high as 86.49.2 months later, and the percentage of medullary cells in spleen was 47.17% and 5.4%, respectively. In bone marrow, medullary cells accounted for 54.27% and 23.5%, respectively, and the percentage of blood cells in peripheral blood was lower, and the percentage of TnB and myeloid cells was 0.726.42% and 2.69%, respectively. The results show that the new liver transplantation method can achieve high efficiency and comprehensive hematopoietic reconstitution. 3. In E8.5 days intrauterine yolk sac transplantation test, 22 healthy newborn rats were obtained. Except for one mouse, the other results were basically below 1%. Eight healthy born mice were obtained by intrauterine transplantation of E12.5 days, 6 of which detected positive expression of human CD45, but the reconstruction rate was lower than that of yolk sac transplantation. In this study, NOD/SCID fetal yolk sac transplantation was performed and 11 healthy offspring were obtained, but the reconstruction rate was less than 1. The results showed that yolk sac transplantation was superior to abdominal transplantation, but the reconstruction rate of intrauterine transplantation was lower than that of other transplantation methods. In addition, the advantage of intrauterine transplantation for hematopoietic reconstitution in immunodeficient mice was not found in this study.
【学位授予单位】：内蒙古大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R-332
【引证文献】
相关期刊论文 前1条
1 魏春山;唐海鸿;童光东;陈孝银;;不同来源肝干细胞的定向分化潜能[J];中国组织工程研究;2012年45期
，本文编号：2413122
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2413122.html
最近更新
教材专著
