2. Academic Validation
  3. SB216763 enhances proliferation and sustains the undifferentiated state of porcine spermatogonial cells in prolonged In vitro culture

SB216763 enhances proliferation and sustains the undifferentiated state of porcine spermatogonial cells in prolonged In vitro culture

  • Theriogenology. 2026 Apr 1:254:117816. doi: 10.1016/j.theriogenology.2026.117816.
Benliang Zhou 1,  Zhen Zhang 1,  Anqin Duan 2,  Jiaxiang Ji 1,  Mengqi Li 1,  Rui Chen 1,  Xiangxing Zhu 3,  Jianghua Shang 4,  Xiaogan Yang 1,  Xingwei Liang 5
Affiliations

Affiliations

  • 1 Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, College of Animal Science & Technology, Guangxi University, Nanning, Guangxi, 530004, China.
  • 2 Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, College of Animal Science & Technology, Guangxi University, Nanning, Guangxi, 530004, China; Guangxi Key Laboratory of Buffalo Genetics, Reproduction and Breeding, Guangxi Buffalo Research Institute, Nanning, 530001, China; Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Ministry of Agriculture and Rural Affairs, Nanning, 530001, China.
  • 3 Gene Editing Technology Center of Guangdong Province, School of Medicine, Foshan University, Foshan, 528225, China.
  • 4 Guangxi Key Laboratory of Buffalo Genetics, Reproduction and Breeding, Guangxi Buffalo Research Institute, Nanning, 530001, China; Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Ministry of Agriculture and Rural Affairs, Nanning, 530001, China.
  • 5 Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, College of Animal Science & Technology, Guangxi University, Nanning, Guangxi, 530004, China. Electronic address: [email protected].
PMID: 41518868 DOI: 10.1016/j.theriogenology.2026.117816
Abstract

Spermatogonial stem cells (SSCs) serve as the foundation of spermatogenesis and play a critical role in livestock fertility and genetic improvement. However, maintaining these cells in long-term culture remains challenging, especially in pigs, where preserving both proliferative capacity and the undifferentiated state has proven difficult. To overcome this limitation, we investigated SB216763 (SB), a selective GSK-3β Inhibitor reported to enhance proliferation in Other stem cell types, for its potential effects on porcine spermatogonial cells (pSCs). In a 9-day concentration screening (0, 1.25, 2.5, and 5 μM SB), SB treatment significantly enhanced the proliferation of pSCs. It upregulated the proliferation-related gene PCNA and the antiapoptotic gene BCL2, while downregulating key apoptosis-related genes, including Bax and Caspase-3. It also promoted the expression of DAZL, UCHL1, NANOS2, and OCT4. Furthermore, over a 55-day period, treatment with 2.5 μM SB consistently supported higher proliferation rates, a larger proportion of DAZL-positive cells, and elevated expression of undifferentiation-associated markers (NANOS2, OCT4) compared with untreated controls at each passage. Overall, our results demonstrate that SB216763 not only promotes the proliferation of pSCs but also helps maintain their germline identity, offering a feasible strategy to improve the long-term in vitro culture of porcine spermatogonial cells.

Keywords

Long-term culture; Porcine spermatogonial cells; Proliferation; SB216763; Undifferentiation.

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