Study Of The Feasibility On That Adipose Tissue-derived Stromal Cells Can Become The Target Cell For Gene Therapy Of Hemophilia A

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-Fâ…§BD) 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 Fâ…§ 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: â‘ Coagulant activity of hFâ…§ was 0.2% when WAV of recombinant plasma and lipofectamine was 0.5:1;â‘¡Coagulant activity of hFâ…§ was 1.6% when W/W of recombinant plasma and lipofectamine was 1:1; â‘¢Coagulant activity of hFâ…§ was 0.4% when W/W of recombinant plasma and lipofectamine was 1.5:1. Coagulant activity of hFâ…§ was 0 when ADSCs were not transducted.Conclusions: Exogenous gene-pRC/RSV-Fâ…§BD can be transfected successfully into adipose tissue-derived stromal cells and be expressed. The ADSCs system may become the perfect target cell.