【摘要】： 造血干細(xì)胞移植是治療血液及免疫系統(tǒng)等疾病的主要手段,通過造血干細(xì)胞移植也可以為多種疾病提供動物模型,尤其是人造血干細(xì)胞移植小鼠建立的人源化血液系統(tǒng)小鼠模型為研究某些無法感染小鼠的人類疾病病毒提供有效途徑。本研究從臍血中分離出人CD34陽性造血干細(xì)胞,分別通過骨髓移植、新生肝移植、宮內(nèi)移植方法建立人源化造血系統(tǒng)小鼠模型。在小鼠移植后1個月至3個月檢測人血細(xì)胞表面抗原CD45。新生肝移植后同時檢測了髓系細(xì)胞表面抗原CD33、T淋巴細(xì)胞表面抗原CD3以及B淋巴細(xì)胞表面抗原CD19。通過人血細(xì)胞表面抗原的表達(dá)檢測出小鼠體內(nèi)人血液系統(tǒng)的重建情況。 1、利用骨髓移植方法共移植12只NOD/SCID小鼠,陰性對照組4只,試驗組8只。試驗組小鼠均取得了人血液細(xì)胞的重建,1個月后檢測外周血人CD45得到46.88%的重建率,3個月后骨髓中人血細(xì)胞重建率達(dá)到96.24%。試驗結(jié)果證明通過骨髓移植人造血干細(xì)胞可以成功促進人造血細(xì)胞在小鼠體內(nèi)的重建。 2、通過新生肝移植方法共移植兩窩16只NOD/SCID仔鼠,陰性對照組4只,試驗組12只。陰性對照小鼠均在2周內(nèi)死亡,未檢測到人血細(xì)胞重建;試驗組小鼠均檢測到人血液細(xì)胞重建,有三只小鼠存活兩個月以上。1個月后檢測外周血人CD45得到26.8%的重建率,2個月后外周血重建率達(dá)到60.94%,骨髓重建率高達(dá)86.49%。2個月后脾臟內(nèi)的人T、B、髓系細(xì)胞分別占4.29%、47.17%和5.4%;骨髓內(nèi)的T、B、髓系細(xì)胞分別占0.25%、54.27%和23.5%;外周血內(nèi)各系血細(xì)胞占的比例較低,T、B、髓系細(xì)胞分別占0.7%、26.42%和2.69%。試驗結(jié)果證明了通過新生肝移植方法能夠得到高效、全面的造血重建。 3、在E8.5d宮內(nèi)卵黃囊移植試驗中,共獲得健康出生仔鼠22只,除一只惺籩亟縭蝕锏?.34%,其他檢測結(jié)果基本處于1%以下。通過E12.5d宮內(nèi)腹腔移植得到健康出生小鼠8只,其中6只檢測到人CD45陽性表達(dá),但重建率總體低于卵黃囊移植得到的重建率。本研究同時進行了NOD/SCID胎鼠卵黃囊移植,獲得健康出生的仔鼠11只,但重建比例亦低于1%。試驗結(jié)果說明卵黃囊移植優(yōu)于腹腔移植,但通過宮內(nèi)移植得到的重建率低于其他移植方式。另外,本研究未發(fā)現(xiàn)免疫缺陷小鼠宮內(nèi)移植對于造血重建的優(yōu)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.
【學(xué)位授予單位】：內(nèi)蒙古大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R-332

【引證文獻(xiàn)】

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1 魏春山;唐海鴻;童光東;陳孝銀;;不同來源肝干細(xì)胞的定向分化潛能[J];中國組織工程研究;2012年45期

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本文編號：2413122