2007 Fiscal Year Final Research Report Summary
ex vivo spermatogenesis from cultured spermatogonial stem cells
| Project/Area Number |
18591783
|
|---|
| 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, Graduate School of Medicine, Department of Urology, Associated professor (50254222)
|
|---|
| Project Period (FY) |
2006 – 2007
|
| Keywords | spermatogenesis / spermatovonial stem cell / culture / micro-insemination |
|---|
| Research Abstract |
1) Establishement and maintenance of mouse spermatogonial stem cell lines:
Three transgenic mice (Tg) lines were used; ubiquitous GFP expressing Tg, Acrosin-GFP Tg expressing GFP at mid-meiosis onward, and Haspin-GFP Tg expressing GFP at haploid stage. Spermatogonial stem cell lines, also called germline stem cells (GS cells) were established from these three Tg mice.
2) Ex vivo spermatogenesis from GS cells:
In order to induce spermatogenesis form GS cells in vitro, GS cells of Acrosin-GFP Tg and Haspin-GFP Tg were cultured in various conditions such as different feeder cells. So far, however, GFP expression, marker of meiosis, was not detected in our experience.
In our trial for ex vivo spermatogenesis, we have found that seminiferous tubules can be reconstituted de novo from dissociated fetal or neonatal testis cells in the subcutis of nude mice. We took advantage of the reconstitution ability of immature testis cells to get GS cells integrated in them. Those GS cells underwent differentiation up to spermatid (haploid) stage which was used for insemination to end up produce healthy pups.
3) Organ culture of testis tissues:
Our experience of cell culture for spermatogenesis was rather discouraging and it becomes evident for us to convert to another strategy. Using acrosin- and haspin-GFP Tg mice testis tissue, 3-14 days old, we evaluated organ culture method. When 7 or more days old pups were used, haspin-GFP become positive in a culture condition. Basic favorable culture condition includes 32℃, 10% fetal bovine serum, vitamins, and so on. Further improvement could be possible in near future.
|
Research Products
(27 results)
-
-
[Journal Article] Identification of transcripts commonly expressed in both hematopoietic and germ-line stem cells.2008
Author(s)
Mizukami T, Kuramitsu M, Takizawa K, Momose H, Masumi A, Naito S, Iwama A, Ogawa T, Noce T, Hamaguchi I, Yamaguchi K.
-
Journal Title
Stem Cell Dev 17
Pages: 67-80
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
-
-
-
[Journal Article] Identification of transcripts commonly expressed in both hematopoietic and germ-line stem cells2008
Author(s)
Mizukami, T, Kuramitsu, M, Takizawa, K, Momose, H, Masumi, A, Naito, S, Iwama, A, Ogawa, T, Noce, T, Hamaguchi, I, Yamaguchi, K
-
Journal Title
Stem Cells Dev 17
Pages: 67-80
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
[Journal Article] Production of Functional Spermatids from Mouse Germline Stem Cells in Ectopically Reconstituted Seminiferous Tubules.2007
Author(s)
Kata K, Watanabe T, Ohsaka K, Hayashi H, Kubota Y, Nagashima Y, Aoki I, Taniguchi H, Noce T, Inoue K, Miki H, Ogonuki N, Tanaka H, Ogura A, Ogawa T.
-
Journal Title
Biology of Reproduction 76
Pages: 211-217
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
-
-
[Journal Article] Production of Functional Spermatids from Mouse Germline Stem Cells in Ectopically Reconstituted Seminiferous Tubules2007
Author(s)
Kita, K, Watanabe, T, Ohsaka, K, Hayashi, H, Kubota, Y, Nagashima, Y, Aoki, I, Taniguchi, H, Noce, T, Inoue, K, Miki, H, Ogonuki, N, Tanaka, H, Ogura, A, Ogawa, T
-
Journal Title
Biol Reprod 76
Pages: 211-217
Description
「研究成果報告書概要(欧文)」より
-
-
-
[Journal Article] Testosterone administration promotes regeneration of chemically impaired spermatogenesis in rats2006
Author(s)
Udagawa, K, Ogawa, T, Watanabe, T, Tamura, Y, Kita, K, Kubota, Y
-
Journal Title
Int J Urol 13(8)
Pages: 1103-1108
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] Differences and similarities in the developmental status of embryo-derived stem cells and primordial germ cells revealed by global expression profiling
Author(s)
Mise, N, Fuchikami, T, Sugimoto, M, Kobayakawa, S, Ike, F, Ogawa, T, Tada, T, Kanaya, S, Noce, T, Abe, K
-
Journal Title
Genes to Cells 2008 (in press)
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
-
-
-
[Presentation] 精子系生殖幹細胞研究の動向2007
Author(s)
小川 毅彦
Organizer
第2回生殖医療エンジニアリング研究会
Place of Presentation
都市センターホテル(東京)
Year and Date
2007-02-25
Description
「研究成果報告書概要(和文)」より
-
-
-
-
