Iranian Journal of Medical Sciences
Iranian Journal of Medical Sciences

Supplementation with Plasma Rich in Growth Factors Improves Short-Term In Vitro Maintenance of Human Fetal Testicular Tissue

Articles InPress

Document Type : Original Article(s)

Authors

Seyyed Amir Moradian 1
Zahra Amirkhani 2, 3
Mansoureh Movahedin 4
Seyyede Nazaninzahra Gharib 5
Hamed Hajipour 1
Kobra Hamdi 6
Behrooz Niknafs 1, 7
Amir Fattahi 1

1 Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran

2 Department of Nursing, School of Nursing, Larestan University of Medical Sciences, Larestan, Iran;

3 Student Research Committee, Larestan University of Medical Sciences, Larestan, Iran

4 Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

5 Department of Biology Education, Shahid Bahonar Campus, Farhangian University, Shiraz, Iran

6 Women’s Reproductive Health Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

7 Department of Anatomy, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

10.30476/ijms.2026.111008.4697
Abstract
Background: Given the need for xeno-free fertility preservation strategies, this study aimed to evaluate plasma rich in growth factors (PRGF) as an alternative to knockout serum replacement (KSR) for short-term in vitro maintenance of human fetal testicular tissue.
Methods: This experimental study was conducted in May 2025 at Al‑Zahra Hospital (Tabriz, Iran). Immature testicular tissue from three 26-week-old human fetal donors was allocated to four culture conditions: 10% KSR (control), 5% PRGF, 10% PRGF, and 20% PRGF. On day 15, 38 tissue fragments were analyzed per culture condition. Fragment-level data were analyzed using a mixed-effects model with culture condition as the fixed effect and fetal donor as the random effect, followed by Tukey’s multiple-comparisons test.
Results: After 15 days of culture, 5% PRGF resulted in better preservation of seminiferous tubules (23.0±2.1%) than 10% KSR and higher PRGF concentrations (P<0.001). It also yielded the highest number of promyelocytic leukemia zinc finger (PLZF)+ (pro)spermatogonia (13.22±1.56 cells/tubule) compared with 10% KSR (11.00±1.65; P=0.0329) and higher PRGF concentrations (P<0.001). Total octamer-binding transcription factor 4/DEAD-box helicase 4 germ cells (OCT4+/DDX4+) were also higher with 5% PRGF than with 10% KSR (13.56±2.60 vs. 8.78±1.39 cells/tubule; P<0.001). In addition, 5% PRGF increased the number of SRY-box transcription factor 9 (SOX9)+ Sertoli cells (P=0.0352), steroidogenic acute regulatory protein (STAR) immunoreactivity (P=0.0121), and Ki‑67+ cells (P=0.0168), while demonstrating lower total Caspase‑3 immunoreactivity (P=0.0109).
Conclusion: PRGF is a promising xeno‑free supplement for short‑term culture of human fetal testicular tissue and merits evaluation in long‑term functional studies.

Highlights

Seyyed Amir Moradian (Google Scholar) 
Amir Fattahi (Google Scholar)

Keywords

Spermatogenesis
Fertility preservation
Organ culture techniques
In vitro techniques
Platelet-rich plasma
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Iranian Journal of Medical Sciences

Articles InPress, Corrected Proof
Available Online from 19 October 2026


  • Received 28 March 2026
  • Revised 21 May 2026
  • Accepted 15 June 2026

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