Journal of Ardabil University of Medical Sciences (Mar 2022)
The Study of Differential and Behavioral Characteristics of Manipulated Human Adipose-Derived Mesenchymal Stem Cells by Lentiviral Vector
Abstract
Background & objectives: Low immunogenicity and targeted migration of mesenchymal stem cells to inflammatory sites have introduced these cells as the best vehicle for the delivery of gene or therapeutic products. Lentiviral vectors can be used as an efficient vehicle for inserting ectopic genes into stem cells. Therefore, the evaluation of the effect of lentiviral vectors on the identity, behavior and functional characteristics of the stem cells can show that vectors are safe tools in cell manipulation and gene therapy. Methods: At first, three vectors of lentivirus were added to HEK-293T cells using calcium phosphate method. After confirming the expression of GFP protein in more than 80% of HEK-293T cells, viral supernatant was collected and concentrated. Human adipose-derived mesenchymal stem cells (hASCs) were transduced with condensed viruses. After the elimination of the non-transduced cells by puromycin, transduced hASCs were evaluated for immunophenotyping and differentiation to adipocyte and osteocyte. Behavioral characteristics such as viability, migration and invasion were analyzed using MTT and transwell methods, respectively. Results: In the current study, there was no significant difference in the expression of CD29, CD34, CD73 and differentiation to adipocyte and osteocyte in the test group when compared to the control group. Moreover, there was no significant difference between two study groups in the behavioral characteristics such as proliferation, invasion and migration. Discussion: The findings of this study declare that lentiviral vectors do not have adverse effects on the identity and functional characteristics of stem cells. Therefore, they can be used in gene manipulation of the target cell.
