Searching of factors facilitate spermatogonial stem cell proliferation (Investigation with transplantation and in vitro analysis)

寻找促进精原干细胞增殖的因素（移植研究和体外分析）

• 批准号: 13671663
• 负责人: OGAWA Takehiko
• 金额: $ 1.54万
• 依托单位: YOKOHAMA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671663/
• 关键词: spermatogenesis; spermatogonia; transplantation; stem cell

1) Fertilization of recipient mice with donor germ cellsPup mice, aged 8 〜 20 days old, were treated with regional radiation to eliminate innate testis germ cells. Spermatogonial transplantation was performed on those mice. Donor spermatogenesis took place in the host testes pervasively at 1 〜 2 months later. Mating with regular females produced donor germ cell derived progeny. This result indicates that spermatogonial stem cells in the pup testis expanded more rapidly than when they were transplanted in the about recipients.2) GnRH analogue effects on spermatogonial cell proliferationWe analyzed proliferation of spermatogonia using transplantation and confocal laser microscopic analysis. In GnRH analogue treated mice, the number of spermatogonia significantly increased compared to those in control mice, indicating that GnRH analogue enhanced proliferation or suppressed apoptosis of the spermatogonia. Another experiment using S1/S1^d mice as recipients showed that the effect of GnRH analogue is independent from SCF signal.3) Culture of spermatogonial stem cellsTestis cells of GFT mice were cultured for 2 to 3 weeks. To confirm the presence of spermatogonial stem cells, they were transplanted to the testes of pup mouse. Several different culture conditions were tested, including feeder cells (STO, 15P-1), coated dishes with lamina or collagen, madrigal matrix, and so on. Among them cultured cells on the madrigal matrix showed they contained spermatogonial stem cells for such periods.

1)受体小鼠与供体生殖细胞的受精用8 ~ 20日龄的Pup小鼠进行局部照射以消除先天睾丸生殖细胞。对这些小鼠进行精原细胞移植。1 ~ 2个月后，供精细胞在宿主睾丸内广泛发生。与正常雌性交配产生供体生殖细胞衍生的后代。这一结果表明，精原干细胞在幼鼠睾丸中的扩增速度比移植到成年鼠睾丸中的快。2）GnRH类似物对精原细胞增殖的影响我们用移植和激光共聚焦显微镜分析了精原细胞的增殖。在GnRH类似物处理的小鼠中，与对照小鼠相比，精原细胞的数量显著增加，表明GnRH类似物促进精原细胞的增殖或抑制精原细胞的凋亡。另一项以S1/S1_d小鼠为受体的实验表明，GnRH类似物的作用不依赖于SCF信号。3）精原干细胞的培养将GFT小鼠睾丸细胞培养2 ~ 3周。为了证实精原干细胞的存在，将它们移植到幼鼠的睾丸中。在饲养层细胞（STO，15 P-1）、胶原包被、基质包被、基质包被等条件下进行了精原干细胞的培养，其中基质包被培养的精原干细胞在此时间段内均为精原干细胞。

项目成果

Ohmura M, Ogawa T, Ono M, Dezawa M, Hosaka M, Kubota Y, Sawada H.: "Increment of Murine Spermatogonial Cell Number by Gonadotropin-Releasing Hormone Analogue Is Independent of Stem Cell Factor-c-kit signal."Biol Reprod.[epub ahead of print]. (2003)

Ohmura M、Okawa T、Ono M、Dezawa M、Hosaka M、Kubota Y、Sawada H.：“促性腺激素释放激素类似物增加小鼠精原细胞数量不依赖于干细胞因子-c-kit 信号。”Biol Reprod

Ohmura M, Ogawa T, et al.: "Increment of Murine Spermatogonial Cell Number by Gonadotropin-Releasing Hormone Analogue Is Independent of Stem Cell Factor -c-kit signal"Biol Reprod. (in press).

Ohmura M、Okawa T 等人：“促性腺激素释放激素类似物增加小鼠精原细胞数量与干细胞因子 -c-kit 信号无关”Biol Reprod。

Ohmura M et al.: "Increment of murine spermatogonial cell number by gonedotropin-releasigng hormone analogue is independent of stem cell factor-c-kit signal"Biology of Reproduction. (in print).

Ohmura M 等人：“促性腺激素释放激素类似物增加小鼠精原细胞数量与干细胞因子 c-kit 信号无关”，《生殖生物学》。

Llolagawa K, Ogawa T, et al.: "GmRH analogue, leuprorclin acetate, promotes regenerarion of rat spermatogenesis after severe chemical damage"Int J Llrol. 8・11. 615-622 (2001)

Llolakawa K、Okawa T 等人：“GmRH 类似物醋酸亮丙酸促进严重化学损伤后大鼠精子发生的再生”Int J Llrol 8・11 (2001)。

Ogawa T et al.: "Expansion of murine spermatogonial stem cells through serial transplantation"Biology of Reproduction. 68. 316-322 (2003)

Okawa T 等人：“通过连续移植扩增小鼠精原干细胞”生殖生物学。