Genetically Modified Adipose Tissue-Derived Stem/Stromal Cells, Using Simian Immunodeficiency Virus-Based Lentiviral Vectors, in the Treatment of Hemophilia B

Abstract

Hemophilia is an X-linked bleeding disorder, and patients with hemophilia are deficient in a biologically active coagulation factor. This study was designed to combine the efficiency of lentiviral vector transduction techniques with murine adipose tissue-derived stem/stromal cells (mADSCs) as a new method to produce secreted human coagulation factor IX (hFIX) and to treat hemophilia B. mADSCs were transduced with simian immunodeficiency virus (SIV)-hFIX lentiviral vector at multiplicities of infection (MOIs) from 1 to 60, and the most effective dose was at an MOI of 10, as determined by hFIX production. hFIX protein secretion persisted over the 28-day experimental period. Cell sheets composed of lentiviral vector-transduced mADSCs were engineered to further enhance the usefulness of these cells for future therapeutic applications in transplantation modalities. These experiments demonstrated that genetically transduced ADSCs may become a valuable cell source for establishing cell-based gene therapies for plasma protein deficiencies, such as hemophilia.

Allogeneic Transplantation of an Adipose-Derived Stem Cell Sheet Combined With Artificial Skin Accelerates Wound Healing in a Rat Wound Model of Type 2 Diabetes and Obesity

“…Rat ASCs (rASCs) were isolated from the inguinal adipose tissue of Lewis rats (20-33 weeks old, male), which was processed according to a previously reported method (23). Briefly, the isolated adipose tissue was enzymatically digested with 0.1% type A collagenase (Roche Diagnostics, Mannheim, Germany) at 37°C for 1 h. The stromal vascular fraction was collected after centrifugation at 700g for 5 min.…”

Gene therapy for hemophilia

“…More recently, studies have begun to explore expression in alternative types of stem cells. Treatment of murine adipose tissue-derived stem/stromal cells (mADSCs) with LV can induce sustained hF.IX expression in vitro , though it remains to be seen whether these cells can engraft into a recipient and provide sustained correction (118). Similarly, menenchymal stem cells (MSCs) derived from human cord blood can be transduced with retroviral or LV vectors to produce F.IX in vitro and in vivo , though efforts are still underway to optimize the matrix for cell growth and F.IX production (119-122).…”

Endoscopic Transplantation of Mesenchymal Stem Cell Sheets in Experimental Colitis in Rats

“…AD-MSC were isolated from the inguinal adipose tissue of SD(CAG-EGFP) transgenic rats, which was processed according to a previously reported method 34 , 35 . Briefly, the isolated adipose tissue was enzymatically digested.…”