Expression Analysis of GDNF/RET Signaling Pathway in Human AD-MSCs Grown in HEK 293 Conditioned Medium (HEK293-CM)
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ORIGINAL PAPER
Expression Analysis of GDNF/RET Signaling Pathway in Human AD-MSCs Grown in HEK 293 Conditioned Medium (HEK293-CM) Zahra Esmaeilizadeh1 Bahar Mohammadi1 Masoumeh Rajabibazl2 Sayyed Mohammad Hossein Ghaderian1 Mir Davood Omrani1 Zahra Fazeli 1 ●
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Received: 1 October 2019 / Accepted: 5 August 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Mesenchymal stem cells have been considered as the suitable source for the repair of kidney lesions. The study and identification of novel approaches could improve the efficiency of these cells in the recovery of kidney. In the present study, the effect of HEK 293 conditioned medium (HEK293-CM) was evaluated on the expression of GDNF/RET signaling pathway and their downstream genes in the human adipose-derived mesenchymal stem cells (AD-MSCs). For this purpose, the human AD-MSCs were cultured in the medium containing HEK293-CM. After the RNA extraction and cDNA synthesis, the expression level of GFRA1, GDNF, SPRY1, ETV4, ETV5, and CRLF1 genes were determined by SYBR Green Real time PCR. The obtained results indicated that the GDNF and GFRA1 expression enhanced in the AD-MSCs following treatment with 10% HEK293-CM-5%FBS as compared to the untreated ADMSCs. These results were consistent with the decreased expression of SPRY1. The significant increased expression of ETV4, ETV5, and CRLF1 genes also showed that HEK293-CM activated the GDNF/RET signaling pathway in the AD-MSCs (P < 0.05). The obtained data suggested that the treatment with HEK293-CM activated the GDNF/RET signaling pathway in the human AD-MSCs. Keywords Mesenchymal stem cells HEK293 Conditioned medium Gene expression GDNF/RET signaling pathway ●
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Introduction The kidney diseases were known to be the common cause of death in the worldwide [1]. Different risk factors were associated with renal dysfunction. They included hypertension, diabetes, and poor life quality as well as the inherited kidney conditions [2–4]. The kidney diseases also increased the risk of developing cardiovascular diseases [5].
Supplementary information The online version of this article (https:// doi.org/10.1007/s12013-020-00936-z) contains supplementary material, which is available to authorized users. * Zahra Fazeli [email protected] 1
Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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There were different options to slow the kidney diseases progression. They contain the administration of angiotensin-converting enzyme inhibitors, bicarbonate, and vitamin D supplementations as well as the control of blood pressure, phosphate, and glucose [6]. The design of novel therapeutic methods has been concentrated on the management of the complications accompanied with kidney diseases, increasing life expectancy in the patients affected by renal dysfunction [7]. In the rec
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