Establishment of Culture System Capable of Inducing Neural Crest Cells from ES Cells

ES细胞诱导神经嵴细胞培养体系的建立

• 批准号: 13680808
• 负责人: KUNISADA Takahiro
• 金额: $ 2.43万
• 依托单位: Gifu 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-13680808/
• 关键词: ES cell; in vitro differentiation; neural crest cell; melanocyte; endothelin 3; retinoic acid; 神経冠細胞; 細胞分化

Mouse embryonic stem (ES) cells are totipotent, primitive, ectodermal-like cells derived from inner cell mass of 3,5 day blastocysts. As an in vitro model for melanocyte differentiation, we have developed the culture system to induce melanocytes from totipotent ES cell line. When undifferentiated ES cells were co-cultured with a bone marrow-derived stromal cell line, a very small but significant number of melanocytes were reproducibly generated. This process was greatly enhanced by addition of dexamethasone to the culture and strictly depended on the c-Kit signaling and endothelin 3 signaling. In this study, we found that caudalizing factor retinoic acid enhanced the efficiency of melanocyte induction in this culture. The early precursors for melanocytes, which proliferate clonally after several passages, were efficiently induced in the culture and their self-renew was stimulated by the addition of endothelin 3 and also by retinoic acid. These results suggests that neural crest cells are induced in this culture system upon corresponding in vivo development.

小鼠胚胎干 (ES) 细胞是全能、原始、外胚层样细胞，源自 3.5 天囊胚的内细胞团。作为黑素细胞分化的体外模型，我们开发了从全能 ES 细胞系诱导黑素细胞的培养系统。当未分化的 ES 细胞与骨髓来源的基质细胞系共培养时，可重复生成极少量但数量显着的黑素细胞。通过向培养物中添加地塞米松，该过程大大增强，并且严格依赖于 c-Kit 信号传导和内皮素 3 信号传导。在这项研究中，我们发现尾化因子视黄酸增强了这种培养物中黑素细胞诱导的效率。黑素细胞的早期前体细胞在传代数次后克隆增殖，在培养物中被有效诱导，并且内皮素3和视黄酸的添加刺激了它们的自我更新。这些结果表明神经嵴细胞在相应的体内发育过程中在该培养系统中被诱导。

项目成果

Wang, K., Gan, L., Kunisada, T., 他3名: "Characterization of the Japanese pufferfish (Takifugu rubripes) T-cell receptor alpha locus reveals a unique genomic organization"Immunogenet.. 53. 31-42 (2001)

Wang, K.、Gan, L.、Kunisada, T. 和其他 3 人：“日本河豚 (Takifugu rubripes) T 细胞受体 α 位点的表征揭示了独特的基因组组织”Immunogenet.. 53. 31-42 ( 2001）

Wang, K., Gan, L., Kunisada, T., Lee, I., Yamagishi, H. and Hood, L.: "Characterization of the Japanese pufferfish (Takifugu rubripes) T-cell receptor alpha locus reveals a unique genomic organization"Immunogenet. 53. 31-42 (2001)

Wang, K.、Gan, L.、Kunisada, T.、Lee, I.、Yamagishi, H. 和 Hood, L.：“日本河豚 (Takifugu rubripes) T 细胞受体 α 基因座的表征揭示了独特的基因组

Wang, K. 他: "Characterization of the Japanese pufferfish(Takifugu rubripes)T-cell receptor alpha locus reveals a unique genomic organization"Immunogenet. 53. 31-42 (2001)

Wang, K. 等人：“日本河豚 (Takifugu rubripes) T 细胞受体 α 基因座的表征揭示了独特的基因组组织”Immunogenet。 53. 31-42 (2001)

Yamane, T., Hayashi, S.I., Kunisada, T.: "Embryonic stem cells as a model to study melanocyte lineage"Methods Mol. Biol.. 185. 261-268 (2002)

Yamane, T.、Hayashi, S.I.、Kunisada, T.：“胚胎干细胞作为研究黑素细胞谱系的模型”方法分子。

Yamane, T., Kunisada, T., Tsukamoto, H., Yamazaki, H., Niwa, H., Takada, S. and Hayashi, S-I.: "Wnt signaling regulates hemopoiesis through stromal cells."J. Immunol. 167. 765-772 (2001)

Yamane, T.、Kunisada, T.、Tsukamoto, H.、Yamazaki, H.、Niwa, H.、Takada, S. 和 Hayashi, S-I.：“Wnt 信号通过基质细胞调节造血。”