Optimizing Electroporation Conditions to ImproveThe Transfection Efficiency of S tem Cells from Apical Papilla

Objective: S tem cells from apical papilla (SCAP) are amongthe mos t widely used s tem cells derived from the biowas tesource. They have been recently considered as a therapeuticcandidate with immunomodulatory potential as well as secretionof a wide variety of neurotrophic factors. But some challengesof genetic manipulation such as the low transfection efficiencyof SCAP are one of the problems that need to be solved.Electroporation which is defined as the process of pore formationin cell membranes via the application of an electric field isa relatively safe and simple method to introduce vectors intothe cells.Materials and Methods: During this s tudy, the cells were maintainedin a normal conditioned medium. In order to electroporation,a single cell suspension was prepared and an aPX۴۶۱vector containing EGFP reporter with different voltages ۱۰۰,۱۵۰, ۲۰۰, ۲۵۰, and ۳۰۰V, double pulse, and pulse width of ۱۰mswere introduced to the SCAPs. ۲۴ hours later, the rate of celldeath and expression of the reporter gene was analyzed with aflow cytometer and fluorescence microscopy.Results: Our results showed that ۲۵۰V provides the highes t efficiencyof SCAPs around ۱۰%.Conclusion: It provided a valuable set-up to improve the transfection efficiency of mesenchymal cells, especially SCAP.

کلیدواژه ها:

Electroporation ، S tem Cells from Apical Papilla ، Transfection Efficiency