Original scientific paper

https://doi.org/10.24099/vet.arhiv.1328

Induced pluripotent stem cell generation using mRNAs: the effect of valproic acid, 5-azacytidine and ascorbic acid

Mahmoud F. Ahmed orcid.org/0000-0002-4654-5909 ; Key Laboratory of Animal Breeding, reproduction and molecular design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, China / Department of Surgery, Anesthesiology and Radiology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt
Ahmed K. El-Sayed ; Key Laboratory of Animal Breeding, reproduction and molecular design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, China / Department of Anatomy and Embryology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt
Eman A. Ahmed ; Department of Pharmacology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt
Yani Zhang ; Key Laboratory of Animal Breeding, reproduction and molecular design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Bichun Li ; Key Laboratory of Animal Breeding, reproduction and molecular design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, China

Abstract

In the bourgeoning fields of tissue engineering and regenerative medicine, induced pluripotent stem cells (iPSCs)
technology with gene therapy are promising candidates for alternative stem cell source and cell transplantation. In this
study, small molecules as anti-oxidant; ascorbic acid (ASA), histone deacetylase inhibitors (HDACi); Valproic acid
(VPA), and DNA methyltransferase inhibitors (DNMTi); 5-Azacytidine (5-AzaC) were examined during the generation
of murine iPSCs using mRNA of Yamanaka factors from mouse embryonic fibroblasts (MEFs). These modulators
were selected based on their well-known effect on the epigenetic status and chromatin modification during early
reprogramming. iPSC generation was performed by using synthesized mRNAs of Yamanaka factors Oct4, Sox2, c-Myc,
and Klf4 (OSCK) as a standard reprogramming strategy. Both morphological changes and the expression level of the
pluripotency markers were examined. 5-AzaC with 1 μM concentration has a slightly toxic effect on the cells, affecting
its proliferation and growing efficiency. In contrast, the use of VPA or ASA led to a two-fold increase in the number of
iPSC colonies. The iPSCs cultured with the addition of VPA or ASA showed a high expression of the tested pluripotency
markers, with a significant increase, more than that of the cells cultured with the addition of 5-AzaC. These findings shed
light on the role of ASA, VPA, and 5-AzaC during murine iPSCs generation using a mRNA reprogramming strategy.

Keywords

iPSCs; 5-azacytidine; valproic acid; ascorbic acid; epigenetics

Hrčak ID:

274814

URI

https://hrcak.srce.hr/274814

Publication date:

24.3.2022.

Article data in other languages: croatian

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