Related Experiment Video
Updated: Mar 31, 2026
Extra Cellular Matrix-Based and Extra Cellular Matrix-Free Generation of Murine Testicular Organoids
Published on: October 7, 2020
Effect of KnockOut serum replacement on germ cell development of immature testis tissue culture
Feng Liu1, Chunhong Cai1, Xin Wu1
1Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
Abstract:
To compare KnockOut serum replacement (KSR) and fetal bovine serum (FBS) for the development of germ cells. Testicular tissues from Sprague-Dawley rats were cultured for 4 weeks in culture media supplemented with FBS or KSR. Tissue area was measured at the beginning and end of the culturing period. Testicular histology, development of the germ cells, and the diameter of seminiferous tubules were analyzed by hematoxylin and eosin staining. After 4 weeks in culture, apoptosis and expression of the stage-specific spermatogenesis marker genes Kit, Sycp3, and Crisp1 were assayed. Tissues cultured in KSR-supplemented media were able to sustain growth and gradually increase seminiferous tubule diameter during the culture period. In addition, spermatogonia, primary spermatocytes, secondary spermatocytes, and round spermatids were observed after 4 weeks in culture, and reverse transcription-PCR confirmed expression of the marker genes. In comparison, tissues cultured in FBS-supplemented media showed dwindling testicular organization, necrotic seminiferous tubules, and expression of Kit, but inconsistent expression of Sycp3 and Crisp1 KnockOut serum replacement outperforms FBS as a growth media supplements for culturing immature spermatogonial tissue culture.
Insights
KnockOut serum replacement (KSR) supports immature rat testicular tissue growth and germ cell development better than fetal bovine serum (FBS). KSR promotes sustained tubule diameter and expression of key spermatogenesis genes, outperforming FBS in this germ cell culture application.
Area of Science:
- Reproductive biology
- Cell culture technology
- Spermatogenesis research
Background:
- Germ cell development and spermatogenesis require specific culture conditions.
- Fetal bovine serum (FBS) is a common supplement, but its efficacy for immature testicular tissue culture is variable.
- Optimizing culture media is crucial for advancing reproductive research and therapies.
Purpose of the Study:
- To compare the efficacy of KnockOut serum replacement (KSR) and FBS as supplements for culturing immature rat testicular tissue.
- To evaluate the impact of KSR versus FBS on tissue viability, germ cell development, and spermatogenesis marker gene expression.
- To determine the optimal serum supplement for maintaining testicular structure and function in vitro.
Main Methods:
- Immature Sprague-Dawley rat testicular tissues were cultured for 4 weeks with either KSR or FBS supplementation.
- Tissue area, seminiferous tubule diameter, and histology were assessed using hematoxylin and eosin staining.
- Apoptosis and expression of stage-specific spermatogenesis markers (Kit, Sycp3, Crisp1) were analyzed via RT-PCR.
Main Results:
- KSR-supplemented cultures maintained tissue integrity and increased seminiferous tubule diameter.
- KSR supported the development of various germ cell types (spermatogonia to round spermatids) and consistent marker gene expression.
- FBS-supplemented cultures showed tissue degradation, necrotic tubules, and inconsistent expression of key spermatogenesis genes.
Conclusions:
- KnockOut serum replacement (KSR) is superior to fetal bovine serum (FBS) for culturing immature rat testicular tissue.
- KSR effectively supports germ cell development and preserves testicular structure in vitro.
- These findings highlight KSR as a preferred supplement for optimizing immature spermatogonial tissue culture in reproductive research.
