Differentiation potency of hematopoietic stem cell line

造血干细胞系的分化能力

• 批准号: 06808074
• 负责人: NAKATSUJI Takako
• 金额: $ 1.28万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06808074/
• 关键词: hematopoietic stem cell; AGM region; differentiation potency; 胚幹細胞; AGM; 分化能; 発生運命; ES細胞; ImmortoMouse; 細胞の分化; 細胞増殖と分化制御; トランスジェニックマウス

We have tried to establish hematopoietic stem cell lines by using a transgenic mouse strain (Immorto Mouse, Charles River Laboratory) harboring SV40 tsA 58 gene. To obtain the most primitive hamatopoietic stem cell population, we dissected out the yolk sac region from embryos at 8 days post coitum (dpc). Such isolated cells were cultured at 33ﾟC in the presence of interferon-gamma to promote transgene expression. They were maintained by subculturing every week for more than 6 months, during which cell cloning was performed. Such established cell lines were tested for their differentiation potency. So far, we obseved appearance of the macrophages and megakaryocytes.As the second source of the early hematopoietic stem cells, we dissected out the ventral trunk region including the paraaortic splanchnopleura from embryos at 10.0 dpc. Recent reports indicate that this region produces the definitive hematopoietic stem cells. Microexplant culture of these tissues produced a cell line that has distinct cell morphology. In subculturing, these cells first produced an epithelia-like cell sheet, but at later stages, they produced hematoocyte-like small round cell population growing on top of the cell sheet. We are currently testing their differentiation potency.

我们尝试用携带SV 40 tsA 58基因的转基因小鼠（Immorto Mouse，Charles River Laboratory）建立造血干细胞系。为了获得最原始的造血干细胞群，我们解剖出卵黄囊区，从胚胎在8天后coconut（dpc）。在干扰素-γ存在下，在33 ℃下培养这种分离的细胞以促进转基因表达。每周传代培养，维持6个月以上，期间进行细胞克隆。测试这些建立的细胞系的分化效力。在胚胎发育10.0dpc时，我们解剖了包括腹主动脉旁内脏胸膜在内的腹主动脉干区域，作为早期造血干细胞的第二来源。最近的报道表明，该区域产生永久性造血干细胞。这些组织的微外植体培养产生了具有不同细胞形态的细胞系。在传代培养中，这些细胞首先产生上皮样细胞片，但在后期阶段，它们产生在细胞片顶部生长的血细胞样小圆细胞群。我们目前正在测试它们的分化能力。

项目成果

中辻憲夫他: "Gene trausfection of mouse primendial germ cells in vitro and analysis of their suruival and growth contmol" Exp.Cell.Res.230巻. 76-83 (1997)

Norio Nakatsuji 等人：“小鼠原始生殖细胞的体外基因转染及其存活和生长控制分析”Exp.Cell.Res.230 (1997)。

中辻憲夫: "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice." Nature. 377. 539-544 (1995)

Norio Nakatsuji：“FAK 缺陷小鼠细胞的细胞运动性降低，粘着斑接触形成增强。” Nature 377. 539-544 (1995)

中辻孝子: "Establishment of cell lines from the yolk sac blood islands of transgenic mouse embryos expressing an immortaligity gene." Cell structure and Function. 発表予定. (1996)

Takako Nakatsuji：“从表达永生基因的转基因小鼠卵黄囊血岛中建立细胞系”（细胞结构和功能）。

中辻孝子他: "Ice・Goby (Shiro-uo),Leucopsarion Petersii,may he a useful matenial for studying Teleostean Embryogenesis" Zoological Science. 14巻・3号. (1997)

Takako Nakatsuji 等人：“Ice Goby (Shiro-uo), Leucopsarion Petersii，可能是研究硬骨动物胚胎发生的有用材料”《动物学科学》第 14 卷，第 3 期。(1997)

Nakatsuji.T., Yamamoto, H., & Nakatsuji, N.: "Establishment of cell line from the yolk sac blood islands and paraaortic splanchnopleura of transgenic mouse embryos harboring an immortalizing gene." Cell Struct, and Funct. 21. 624 (1997)

Nakatsuji.T.、山本 H.、