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Culture bovine prospermatogonia with 2i medium
Huan Cai1, Yu Jiang1, Sheng Zhang2
1College of Veterinary Medicine, Jilin University, Changchun, China.
Andrologia
|March 25, 2021
Summary
Cryopreservation and expansion of bovine prospermatogonia are challenging. A novel 2i medium effectively supports cultivation of these cells from cryopreserved testes, improving germplasm preservation.
Area of Science:
- Reproductive Biology
- Stem Cell Biology
- Cryobiology
Background:
- Germplasm cryopreservation and expansion of bovine prospermatogonia (precursors to sperm) are crucial for cattle breeding but technically difficult.
- Inhibitors of Mek1/2 and Gsk3β (2i) are known to maintain pluripotency in stem cells.
- The efficacy of 2i-based medium for cultivating bovine prospermatogonia from cryopreserved testicular tissue requires investigation.
Purpose of the Study:
- To evaluate the effects of a 2i-based medium on the cultivation and expansion of bovine prospermatogonia derived from cryopreserved testicular tissues.
- To assess the impact of 2i medium on the expression of key pluripotency and differentiation markers, apoptosis, and epigenetic modifications in cultured prospermatogonia.
Main Methods:
- Newborn bull testicular tissues were cryopreserved.
- Prospermatogonia were enriched using differential plating and Percoll-based centrifugation.
- Cultivation was performed in a 2i-based medium, and effects on cell proliferation, gene/protein expression, apoptosis, and histone methylation were analyzed.
Main Results:
- Cryopreserved testicular tissues retained viable prospermatogonia expressing key markers (PLZF, GFRα-1, Oct4/Pouf5, Sox2, Nanog).
- The 2i medium significantly enhanced prospermatogonia proliferation, boosted pluripotency and anti-apoptotic gene expression, and suppressed differentiation markers.
- 2i medium reduced apoptosis, decreased histone H3 lysine (K9) trimethylation (H3K9me3), and lowered the expression of its methylase (Suv39h1/2).
Conclusions:
- The 2i-based medium effectively supports the cultivation and expansion of bovine prospermatogonia from cryopreserved testes.
- This improvement is mediated by inhibiting Suv39h1/2-mediated H3K9me3 through Mek1/2 and Gsk3β signaling pathways.
- The findings demonstrate successful cryopreservation and expansion of bovine germplasm, offering a promising avenue for conservation and breeding.
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