| Project/Area Number |
17591706
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | St.Marianna University School of Medicine |
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Principal Investigator |
IWAMOTO Teruaki St.Marianna University School of Medicine, School of Med., Dept. of Urol., Professor (60046117)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Yoko Nagasaki Univ., Dept. of Cell Biol. and Histochem., Assistant Professor (50398963)
YOSHIIKE Miki St. Marianna University School of Medicine, Dept. of Urol., Research Technician (60398964)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2006: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | spermatogenesis / lamina propria / glycoprotein / development-differentiation / cell line |
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| Research Abstract |
Thickened lamina propria in seminiferous tubules of the testis is unique morphological feature of human testis showing deteriorated spermatogenesis. Previously, we showed that inner layer of thickened lamina propria had glycoproteins recognized by PNA-lectin and showed the property to bind progesterone. To clarify the role of lamina propria in spermatogenesis, we performed further analysis to identify products. Testicular samples from a patient showing almost normal spermatogenesis and a patient showing deteriorated spermatogenesis were homogenized and solubilized. The solubilized proteins were analyzed by 2-D gel electrophoresis. The blotting membrane was pre-incubated in the solution with or without progesterone, and then the positive spots were screened by PNA-lectin affinity. This screening method could select the spots having both PNA-lectin affinity and progesterone binding affinity. However, we need to know the optimal condition for this screening method to determine the concerned glycoproteins in lamina propria because there are still several spots. As another projects, we tried to establish the immortalized cell line derived from human testis to investigate the synthesis of glycoproteins under cellular level. Co-culture of the primary cultured human testicular cells and mouse Sertoli cell lines (A31C4) allowed us to maintain the human spermatogonia and Sertoli cells. Furthermore, the Sertoli cells from adult human testis could proliferate under the co-culture condition. Further steps for separation of Sertoli cells derived from human and mouse are needed, however, we believe that this co-culture system will contribute to the establishment of cell line from human testicular cells and the induction of differentiation of human spermatogonia in vitro.
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