Functional analysis and culture of human spermatogonial stem cells
| Project/Area Number |
19K22512
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 48:Biomedical structure and function and related fields
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Project Period (FY) |
2019-06-28 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2021: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2020: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | 精子形成 / 精子幹細胞 / ヒト / 幹細胞 / 培養 |
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| Outline of Research at the Start |
げっ歯類の精子幹細胞研究の進展に伴い、海外ではヒトの精子幹細胞研究に興味が高まっている(Nat Med 2013;19:958)。しかしながら、ヒトの精子幹細胞についてはこれまでほとんど研究されておらず、その同定すら困難である。幹細胞は機能的にしか同定できないため、ヒトの精子幹細胞の機能的アッセイの同定は重要な課題である。そこで本研究ではこの問題を解決するためにヒト精子幹細胞のin vivoもしくはin itroの機能的アッセイ系の確立を目指すと共にヒト精子幹細胞の試験管内での自己複製の再現を試みる。
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| Outline of Final Research Achievements |
To set up a culture system for human spermatogonial stem cells (SSCs), we established a method for enzymatic digestion of human testis cells. Based on the gene expression patterns in mouse SSCs, we screened monoclonal antibodies that can react with human SSCs. Through this screening, we found that human spermatogonial stem cells express Epha2 on their surface. The frequency of cells expressing Epha2 in the testis is significantly lower than those previously reported for human SSCs. This purification protocol was useful for starting human SSC cultures without the contamination of testicular somatic cells. By adjusting the culture conditions, we were able to derive two candidate SSC cultures. However, we still have not been able to collect these cells for functional analysis because they died upon dissociation into single cells by trypsin. We are currently taking different approaches to determine their SSC activity.
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| Academic Significance and Societal Importance of the Research Achievements |
癌治療成績の向上に伴い、小児の約7-8割の患者が生存し、20代の若者250人に一人が癌治療の生存者である。ところが、治療の副作用として半分程度の患者が不妊となっている。ヒト精子幹細胞の培養が成功すれば、この細胞をがん治療前の患者さんの精巣から回収し、試験管内で増幅したのちに治療後の精巣に移植するすることで妊孕性を回復することができると期待されている。今回の研究によりヒト精子幹細胞の培養条件が改善し、近い将来ヒトへの応用も可能になると期待できる。
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Report
(4 results)
Research Products
(17 results)
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[Journal Article] ROS amplification drives mouse spermatogonial stem cell self-renewal.2019
Author(s)
Morimoto H, Kanatsu-Shinohara M, Ogonuki N, Kamimura S, Ogura A, Yabe-Nishimura C, Mori Y, Morimoto T, Watanabe S, Otsu K, Yamamoto T, *Shinohara T.
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Journal Title
Life Sci Alliance
Volume: 2
Issue: 2
Pages: 2-2
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Aging of spermatogonial stem cells by Jnk-mediated glycolysis activation.2019
Author(s)
Kanatsu-Shinohara, M., Yamamoto, T., Toh, H., Kazuki, Y., Kazuki, K., Imoto, J., Ikeo, K., Oshima, M., Shirahige, K., Iwama, A., Nabeshima, Y., Sasaki, H. and Shinohara, T.
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Journal Title
PNAS
Volume: 116
Issue: 33
Pages: 16404-16409
DOI
Related Report
Peer Reviewed
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