Searching for culture condition optimal for proliferation of spermatogonial stem cells
| Project/Area Number |
15591702
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Urology
|
|---|
| Research Institution | YOKOHAMA CITY UNIVERSITY |
|---|
Principal Investigator |
OGAWA Takehiko Yokohama City University, School of Medicine, Associate Professor, 医学部, 講師 (50254222)
|
|---|
| Project Period (FY) |
2003 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,100,000 (Direct Cost: ¥1,100,000)
|
|---|
| Keywords | spermatogenesis / stem cell / cell culture |
|---|
| Research Abstract |
Culturing of Spermatogonial Stem Cells
In April, 2003, it was reported that male germline stem cells, collected from neonatal mouse testes (strains of DBA/2 and ICR), were cultured for long period on feeder layer of mouse embryonic fibroblasts with 4 growth factors (EGF, bFGF, LIF, GDNF). We immediately repeated and confirmed results of the report and progressed it one step beyond by establishing germline stem cells (GS cells) from not only neonates but also from adult mice. The GS cells expanded exponentially in vitro. They produce sperm when transplanted recipient mouse testes. They did not produce tumor nor showed abnormal proliferation in the body of recipient mice. The GS cells was re-derived from recipient mouse testes, which showed that GS cells in vitro and spermatogonial stem cells in the testis were identical or mutually convertible.
Fertilization of recipient mice with spermatogonial transplantation
In order to make recipient mice fertile after spermatogonial transplantation, immature mice (10-15 days old) were treated with irradiation before transplantation. The aim of irradiation was to make empty niche in the testis by removing endogenous germ cells for efficient colonization of donor stem cells. The dose of irradiation turned out to be optimal at around 12Gy for that purpose. One to Three days after the irradiation, the recipient mice received spermatogonial transplantation. Two months after the transplantation onward, the recipient mice were mated with females to test their fertility. It was confirmed that mice became fertile fathered pups of donor germ cell origin at high rate (about 70%). When we used GS cells, the fertility rate of recipient mice became higher, around 80%. The important factors for making recipient fertile with donor germ cells therefore include age of recipient, stem cell concentration of donor cell suspension.
|
Report
(3 results)
Research Products
(14 results)
-
-
-
-
[Journal Article] Oligo-astheno-teratozoospermia in mice lacking Cnot7, a regulator of retinoid X receptor beta2004
Author(s)
Nakamura, T., Yao, R., Ogawa, T., Suzuki, T., Ito, C., Tsunekawa, N., Ajima, R., Miyasaka, T., Yoshida, Y., Toshimori, K., Noce, T., Yamamoto, T., Noda, T.
-
Journal Title
Nature Genetics 36
Pages: 528-533
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Journal Article] Derivation and Morphological Characterization of Mouse Spermatogonial Stem Cell Lines2004
Author(s)
Ogawa, T., Ohmura, M., Tamura, Y., Kita, K., Ohbo, K., Suda, T., Kubota, Y.
-
Journal Title
Arch. Histol. Cytol. 67
Pages: 297-306
NAID
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Journal Article] Abnormal sperm morphology caused by defects in Sertoli cells of Cnot7 knockout mice2004
Author(s)
Ogawa, T., Ito, C., Nakamura, T., Tamura, Y., Yamamoto, T., Noda, T., Kubota, Y., Toshimori, K.
-
Journal Title
Arch. Histol. Cytol. 67
Pages: 307-314
NAID
Description
「研究成果報告書概要(欧文)」より
Related Report
-
-
-
-
-
-
[Journal Article] Increment of murine spermatogonial cell number by Gonadotropin-releasing hormone analogue is independent of stem cell factor-c-kit signal2003
Author(s)
Ohmura, M., Ogawa, T., Ono, M., Dezawa, M., Hosaka, M., Kubota, Y., Sawada, H.
-
Journal Title
Biol Reprod. 68
Pages: 2304-2313
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Journal Article] Identification and characterization of stem cells in prepubertal spermatogenesis in mice2003
Author(s)
Ohbo, K., Yoshida, S., Ohmura, M., Ohneda, O., Ogawa, T., Tsuchiya, H., Kuwana, T., Kehler, J., Abe, K., Scholer, H.R., Suda, T.
-
Journal Title
Dev Biol. 258
Pages: 209-225
Description
「研究成果報告書概要(欧文)」より
Related Report
-
