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Study of the Feasibility on That Adipose Tissue-derived Stromal Cells Can Become the Target Cell for Gene Therapy of Hemophilia A

Author: LiYinHua
Tutor: HuangXingYuan
School: Wuhan University
Course: Pediatrics
Keywords: hemophilia A stromal cells Gene expression
CLC: R554
Type: Master's thesis
Year: 2005
Downloads: 54
Quote: 0
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Abstract


Objective: To explore the feasibility of adipose tissue-derived stromal cells (ADSCs) used for gene therapy of hemophilia A.Methods: The experiment has finished in the lab of pathophysiology of Medical College of Wuhan University from 2004.1 to 2004.6. At first, recombinant plasma (pRC/RSV-FⅧBD) were increased、 purified and identified. ADSCs were separated from celiac and postkidney fat tissue. ADSCs were transduced with a recombinant plasma and lipofectamine at different ratio (W/W0.5:1, 1:1, 1.5:1) when they arrived at jion of 80%. Coagulant activity of human FⅧ and transduction efficiencies of ADSCs were measured by colorimetry and semi-quantitative PCR, respectively.Results: Transduction efficiencies was higher 5% when W/W was 1.5:1 than when WAV was 0.5:1. Transduction efficiencies was higher 50% when W/W was 1:1 than when W/W was 0.5:1. Coagulant activity was measured 36h after ADSCs were transducted. Result showed: ①Coagulant activity of hFⅧ was 0.2% when WAV of recombinant plasma and lipofectamine was 0.5:1;②Coagulant activity of hFⅧ was 1.6% when W/W of recombinant plasma and lipofectamine was 1:1; ③Coagulant activity of hFⅧ was 0.4% when W/W of recombinant plasma and lipofectamine was 1.5:1. Coagulant activity of hFⅧ was 0 when ADSCs were not transducted.Conclusions: Exogenous gene-pRC/RSV-FⅧBD can be transfected successfully into adipose tissue-derived stromal cells and be expressed. The ADSCs system may become the perfect target cell.

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CLC: > Medicine, health > Internal Medicine > Blood and lymphatic system diseases > Blood diseases > Hemorrhagic disease
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