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Experimental Study on Immune Tolerance in Orthotopic Allogenic Liver Graft Induced by Mesenchymal Stem Cell Transplantation in Rats

Author: HuangXiaoJin
Tutor: WangXiaoMin
School: Fujian Medical
Course: Surgery
Keywords: Rat Liver Transplant Bone marrow mesenchymal stem cells Cell transplantation Immune tolerance
CLC: R657.3
Type: Master's thesis
Year: 2008
Downloads: 152
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Abstract


Objective: To observe the different treatment factors on rat survival after liver transplantation in rats explore the effect of bone marrow mesenchymal stem cell transplantation combined with Asian-dose FK506 after liver transplantation immune tolerance induced by the feasibility and possible mechanisms. Materials and Methods: First, to establish a stable and reliable animal model of orthotopic liver transplantation. 160 male SD rats were divided into groups of donor and acceptor groups. Using a modified \From rat bone marrow, separation, purification, in vitro amplification of bone marrow MSC to the third generation, parallel CFDA tag. MSC-labeled rat portal vein injection, 7 d after the serial sections, DAPI staining and observed under a fluorescence microscope rat MSC homing situation. Inbred female Lewis rats and female BN rats were 64, LEWIS as donor (n = 32), BN as recipients (n = 32), production of orthotopic liver transplantation rat model 32, is divided into 4 groups. Group A: receptor liver transplant after portal vein injection of 1 mL normal saline; B Group: receptor liver transplant after portal vein injection of 2 × 10 6 MSCs; C Group: receptor liver transplantation after portal vein injection of 1 mL normal saline, given therapeutic doses of FK506 (0.5 mg / kg.d -1 ); D Group: receptor liver transplant after portal vein injection of 2 × 10 6 MSC, while giving the sub-therapeutic dose of FK506 (0.125 mg / kg.d -1 ). Animals were observed throughout the body, recording the survival time after l4 d monitor serum glutamic alanine aminotransferase (ALT), total bilirubin (TBIL) levels, as well as the pathological changes of liver transplantation; PCR semi-quantitative detection of blood SRY ; flow cytometry BN rat spleen HIS19 antibody detection, evaluation chimera formation. Results: The MSC transplantation and sub-therapeutic dose of FK506 group survival time (52.8 ± 4.1) d was significantly longer than saline control group (12.5 ± 0.8) d and pure MSC injection group (12.7 ± 1.2) d, and the immune inhibitor group (47.8 ± 3.8) d rather, the pathological score based programs are more Banff saline control group [severe (Ⅲ level)] and pure MSC injection group [moderate (Ⅱ grade)] was significantly lower, as mild (Ⅰ level). Joint MSC transplantation and sub-therapeutic dose of FK506 group receptor positive cells in the spleen cells HIS19 content of (13.0 ± 2.5)%, compared with the saline control group (0.4 ± 0.5)%, pure MSCs injection group (0.5 ± 0.3)% and treatment FK506 dose injection group (1.4 ± 0.9)% was significantly increased (P lt; 0.01). In addition, the joint MSC transplantation group and sub-therapeutic dose of FK506, FK506 dose injections of serum ALT levels were (49.48 ± 9.70) U / L and (54.3 ± 11) U / L, compared with the saline control group [(886.81 ± 90.3) U / L] and pure MSC injection group [(869.7 ± 109) U / L] was significantly lower (P lt; 0.01); Joint MSC transplantation and sub-therapeutic dose of FK506 group, dose FK506 injection group TBIL concentrations were (13.52 ± 2.28) μmol / L and (13.35 ± 2.05) μmol / L, compared with the saline control group [(178.72 ± 35.81) μmol / L] and pure MSC injection group [(179.92 ± 37.67) μmol / L] was also significantly lower (P lt; 0.01 or P lt; 0.05). Conclusion: Bone marrow mesenchymal stem cell transplantation can reduce allogeneic liver transplant acute rejection, combined with low dose FK506 can induce the production of micro-chimerism and prolong survival time of transplanted rats, inducing immune tolerance after liver transplantation.

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