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Application of Platelet-Rich Plasma (Prp) Improves Self-Renewal of Human Spermatogonial Stem Cells in Two-Dimensional and Three-Dimensional Culture Systems Publisher Pubmed



Khadivi F1 ; Koruji M2 ; Akbari M1 ; Jabari A3 ; Talebi A4 ; Ashouri Movassagh S5 ; Panahi Boroujeni A6 ; Feizollahi N1 ; Nikmahzar A1 ; Pourahmadi M7 ; Abbasi M1
Authors
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Authors Affiliations
  1. 1. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Stem Cell and Regenerative Medicine Research Center & Department of Anatomy, Iran University of Medical Sciences, Tehran, Iran
  3. 3. Department of Obstetrics and Gynecology, Moluod infertility center, Zahedan University of Medical Sciences, Zahedan, Iran
  4. 4. School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
  5. 5. Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran
  6. 6. Department of Anesthesiology, Shahrekord University of Medical Sciences, Shahrekord, Iran
  7. 7. Department of Anatomy, Jahrom University of Medical Sciences, Jahrom, Iran

Source: Acta Histochemica Published:2020


Abstract

Spermatogonial stem cells (SSCs) are very sensitive to chemotherapy and radiotherapy, so male infertility is a great challenge for prepubertal cancer survivors. Cryoconservation of testicular cells before cancer treatment can preserve SSCs from treatment side effects. Different two-dimensional (2D) and three-dimensional (3D) culture systems of SSCs have been used in many species as a useful technique to in vitro spermatogenesis. We evaluated the proliferation of SSCs in 2D and 3D culture systems of platelet-rich plasma (PRP). testicular cells of four brain-dead patients cultivated in 2D pre-culture system, characterization of SSCs performed by RT-PCR, flow cytometry, immunocytochemistry and their functionality assessed by xenotransplantation to azoospermia mice. PRP prepared and dosimetry carried out to determine the optimized dose of PRP. After preparation of PRP scaffold, cytotoxic and histological evaluation performed and SSCs cultivated into three groups: control, 2D culture by optimized dose of PRP and PRP scaffold. The diameter and number of colonies measured and relative expression of GFRa1 and c-KIT evaluated by real-time PCR. Results indicated the expression of PLZF, VASA, OCT4, GFRa1 and vimentin in colonies after 2D pre-culture, xenotransplantation demonstrated proliferated SSCs have proper functionality to homing in mouse testes. The relative expression of c-KIT showed a significant increase as compared to the control group (*: p < 0.05) in PRP- 2D group, expression of GFRa1 and c-KIT in PRP scaffold group revealed a significant increase as compared to other groups (***: p < 0.001). The number and diameter of colonies in the PRP-2D group showed a considerable increase (p < 0.01) as compared to the control group. In PRP- scaffold group, a significant increase (p < 0.01) was seen only in the number of colonies related to the control group. Our results suggested that PRP scaffold can reconstruct a suitable structure to the in vitro proliferation of SSCs. © 2020 Elsevier GmbH
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