期刊
ANNALS OF BIOMEDICAL ENGINEERING
卷 45, 期 7, 页码 1783-1794出版社
SPRINGER
DOI: 10.1007/s10439-017-1847-z
关键词
In vitro spermatogenesis; Germ cell; DDX4; CREM; KSR
资金
- Jeanssons Foundation
- Sallskapet Barnavard in Stockholm
- Swedish Research Council/Academy of Finland
- Emil and Wera Cornells Foundation
- Stiftelsen Sigurd och Elsa Goljes Minne
- Kronprinsessan Lovisas Forening For Barnasjukvard/Stiftelsen Axel Tielmans Minnesfond
- Samariten Foundation
- Sigrid Juselius Foundations
- Turku University Hospital special governmental grant
- Swedish Childhood Cancer Foundation
- Stockholm County Council
- Karolinska Institutet
- [EU-FP7-PEOPLE-2013-ITN 69-007]
Finding robust culture conditions for in vitro maturation (IVM) of male germ cells is still a challenge. Recently, a testis organ culture method, using Knockout Serum Replacement (KSR), was suggested as a promising approach. However, the efficiency of that model is still not optimal. Hence, we have tried to establish the culture conditions in two laboratories, and to improve the reliability of the culture system to generate mature germ cells. Male mice at three days of age were sacrificed. Testes were cut into small pieces which were cultured atop agarose stands, using Minimum Essential Medium alpha supplemented with different supplements; melatonin, Glutamax, and different concentrations of KSR. The results showed that the duration of culture beyond 18 days had an impact on the number of differentiated germ cells. Supplementation with melatonin and Glutamax revealed a positive influence on the efficiency of male germ cell differentiation in vitro. Furthermore, the results confirmed that KSR had a positive effect on germ cell maturation and testosterone production, with a concentration of at least 10%. In conclusion, this study emphasizes the beneficial role of at least 10% KSR in the IVM of germ cells.
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