Maryam Kaviani1, Massood Ezzatabadipour2, Seyed Noureddin Nematollahi-Mahani3, Parvin Salehinejad4, Mozhgan Mohammadi5, Seyed Mehdi Kalantar6, Batool Motamedi4. 1. Department of Anatomical Sciences, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran. 2. Department of Anatomical Sciences, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; Physiology Research Centre, Kerman University of Medical Sciences, Kerman, Iran; Neuroscience Research Centre, Kerman University of Medical Sciences, Kerman, Iran. Electronic address: m_ezatabadi@kmu.ac.ir. 3. Department of Anatomical Sciences, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; Afzal Research Institute (NGO), Kerman, Iran; Neuroscience Research Centre, Kerman University of Medical Sciences, Kerman, Iran. 4. Department of Midwifery, Kerman University of Medical Sciences, Kerman, Iran. 5. Physiology Research Centre, Kerman University of Medical Sciences, Kerman, Iran; Department of Microbiology, Virology and Immunology, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran. 6. Reproductive and Genetic Unit, Research and Clinical Centre for Infertility, Yazd University of Medical Sciences, Yazd, Iran.
Abstract
BACKGROUND AIMS: Vitrification as an advanced cryopreservation method is recommended for cell storage toward future applications. The purpose of this report was to appraise whether gametogenic potential of these cells is altered by vitrification. METHODS: A two steps method was applied for hUCM cells vitrification. An n-hUCM group of hUCM cells served as control. In order to differentiation of hUCM cells into male germ cells, the cells were induced by retinoic acid, testosterone and testicular-cell-conditioned medium. To evaluate induced hUCM cells toward germ cells, we used immunocytochemistry and karyotyping methods. RESULTS: v-hUCM cells similar to n-hUCM cells formed flat cells after gametogenic induction, and showed protein expression of germ-cell-specific markers DAZL, VASA (DDX4) and SCP3. Karyotyping pattern remained unchanged in the either groups. CONCLUSIONS: The analysis of these results demonstrates that vitrification does not alter differentiation potential of hUCMs to male germ like cells. These results may set an in vitro pattern to study germ-cell formation from hUCM cells and also as a potential source of sperms for male infertility.
BACKGROUND AIMS: Vitrification as an advanced cryopreservation method is recommended for cell storage toward future applications. The purpose of this report was to appraise whether gametogenic potential of these cells is altered by vitrification. METHODS: A two steps method was applied for hUCM cells vitrification. An n-hUCM group of hUCM cells served as control. In order to differentiation of hUCM cells into male germ cells, the cells were induced by retinoic acid, testosterone and testicular-cell-conditioned medium. To evaluate induced hUCM cells toward germ cells, we used immunocytochemistry and karyotyping methods. RESULTS: v-hUCM cells similar to n-hUCM cells formed flat cells after gametogenic induction, and showed protein expression of germ-cell-specific markers DAZL, VASA (DDX4) and SCP3. Karyotyping pattern remained unchanged in the either groups. CONCLUSIONS: The analysis of these results demonstrates that vitrification does not alter differentiation potential of hUCMs to male germ like cells. These results may set an in vitro pattern to study germ-cell formation from hUCM cells and also as a potential source of sperms for male infertility.
Authors: Gina D Kusuma; Mehri Barabadi; Jean L Tan; David A V Morton; Jessica E Frith; Rebecca Lim Journal: Front Pharmacol Date: 2018-10-29 Impact factor: 5.810