Abstract
Mammals have a limited capacity to regenerate their tissues and organs. One of the mechanisms associated with natural regeneration is dedifferentiation. Several small molecules such as vitamin C and growth factors could improve reprogramming efficiency. In this study, the NTERA2-D1 (NT2) cells were induced towards differentiation (NT2-RA) with 10-5 M retinoic acid (RA) for three days and then subjected to various amounts of vitreous humor (VH). Results show that the growth rate of these cells was reduced, while this rate was partly restored upon treatment with VH (NT2-RA-VH). Cell cycle analysis with PI method also showed that the numbers of cells at the S phase of the cell cycle in these cells were increased. The levels of SSEA3 and TRA-1-81 antigens in NT2-RA were dropped but they increased in NT2- RA-VH to a level similar to the NT2 cells. The level of SSEA1 had an opposite pattern. Expression of OCT4 gene dropped after RA treatment, but it was recovered in NT2-RA-VH cells. In conclusion, we suggest VH as a potent mixture for improving the cellular reprogramming leading to dedifferentiation.
Highlights
Mammals have a limited capacity to regenerate their tissues
Embryonic stem (ES) cells, embryonal carcinoma (EC) cells and embryonic germ (EG) cells can induce nuclear reprogramming after their hybridization with somatic cells (Jaenisch and Hochedlinger, 2015; Pfeiffer et al, 2013; Wang et al, 2016; Xu et al, 2016)
The NTERA2 cells were grown in Dulbecco’s Modified Eagles Medium (DMEM) supplied with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin
Summary
Mammals have a limited capacity to regenerate their tissues. Tissue regeneration can be occurred either through the activation of somatic stem cells, located in a niche or by inducing proliferation of the differentiated cells. Takahashi and Yamanaka in 2006 (Takahashi and Yamanaka, 2006) showed that over-expression of 4 genes (Oct, Sox, c-Myc, and Klf4) in mouse embryonic fibroblasts (MEFs), changed the properties of the cells in favor of embryonic. Dedifferentiation involves a terminally differentiated cell reverting back to a less differentiated stage from within its own lineage that allows the cell to proliferate again before redifferentiation This process would lead to replacement of the lost cells. This study sought to investigate the impact of the rabbit VH on proliferation, cell cycling, and self-renewal specific markers’ expression in the model cells of NTERA2
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