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04.03.2019 | Original Article | Ausgabe 4/2019

Comparative Clinical Pathology 4/2019

Comparison of transduction efficiency among various cell types by a lentivector containing CMV promoter

Zeitschrift:
Comparative Clinical Pathology > Ausgabe 4/2019
Autoren:
Hori Ghaneialvar, Abbas Sahebghadam Lotfi, Sareh Arjmand, Leila Soltani, Azra Kenarkoohi, Mohammad Mahdi Zangeneh
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Abstract

Due to their unique biological characteristics, mesenchymal stem cells (MSCs) are useful in developing cell-based therapies, tissue regeneration, and many other applications of cellular and molecular genetics. Use of lentiviral system allows the integration of specific nucleic acid sequence into a target cell genome near an active transcription site in such a way that it is stable and heritable. The aim of the present study was to evaluate the gene transduction efficiency on the cells of different species using lentivirus along with cytomegalovirus in a lentiviral shuttle vector. In this study, the third-passage isolated MSCs from four different species, including ovine and goat cardiac fibroblasts, HEK293T, and L929 cell line were transduced by concentrated virus particles. Then, transduction percentage was detected by flow cytometry. The findings showed different transduction efficiencies for various cells of one species or other species, indicating that differentiated and undifferentiated cells had different transduction percentages. Different degrees of transduction observed in differentiated and undifferentiated cells might be the result of various lentiviral receptors on the plasma membrane, site of integration, and activity of CMV promoter. Further, other regulatory factors involved in cell transduction and target gene expression can cause different percentages of transduction.

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