Abstract Number: PB1093
Meeting: ISTH 2020 Congress
Theme: Hemophilia and Rare Bleeding Disorders » Hemophilia Gene Therapy
Background: Adipose stem cells have many advantages,such as easy to obtain,low immunogenicity,transformed in multiple directions,and can be used as one of the target cells for hemophilia B gene therapy.
Aims: To explore the possibility of successful transfection of adipose-derived stem cells (ADSCs) as target cells for hemophilia B (HB) gene therapy by recombinant adenoviral vector containing human coagulation factor Ⅸ gene (FⅨ).
Methods: ADSCs were isolated and cultured. A recombinant adenovirus vector carrying FⅨ was transfected into ADSCs: group A:(DMEM was used instead of adenovirus to transfect ADSCs),group B:(Ad-GFP was used to transfect ADSCs), and group C: (Ad-hFIX was used to transfect ADSCs),group D:(one generation after passage of Ad-hFIX was used to transfect ADSCs ),group E:(two generations after passage of Ad-hFIX was used to transfect ADSCs).The expression of hFIX gene was detected by RT-PCR. The expression of hFIX protein in ADSCs was detected by Western blot. The hFIX protein amount in the transfected ADSCs was measured by ELISA, the coagulation activity in culture supernatant was measured by one-stage method.
Results: ADSCs were successfully extracted. the expression of hFⅨantigen, gene and protein was detected in ADSCs after transfection of recombinant adenovirus vector, and the expression of hFⅨantigen, gene and protein was able to be stably transfected to the third generation adscs. compared with the blank control group, the difference was statistically significant (p < 0.05).
Conclusions: 1.Adenovirus carrying hFIX gene can effectively transfect ADSCs. 2.ADSCs modified by hFIX gene can secrete hFIX protein with coagulation activity.3.Recombinant adenovirus-transfected ADSCs were cultured after passage and still express hFIX. And ADSCs could serve as vector cells for hemophilia B gene therapy.
[Adenovirus mediates expression of human coagulation factor IX gene in adipose-derived mesenchymal stem cells]
To cite this abstract in AMA style:
Yan ZY, Wang X, Li J, Xie Y, Wang L, Zhu Y, He H, Sun Q. Adenovirus Mediates Expression of Human Coagulation Factor IX Gene in Adipose-Derived Mesenchymal Stem Cells [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/adenovirus-mediates-expression-of-human-coagulation-factor-ix-gene-in-adipose-derived-mesenchymal-stem-cells/. Accessed September 29, 2023.« Back to ISTH 2020 Congress
ISTH Congress Abstracts - https://abstracts.isth.org/abstract/adenovirus-mediates-expression-of-human-coagulation-factor-ix-gene-in-adipose-derived-mesenchymal-stem-cells/