Analysis of the mechanism for organogenesis in mouse embryo and its application to therapeutic transplantation to human fetus

小鼠胚胎器官发生机制分析及其在人类胎儿治疗性移植中的应用

• 批准号: 13470157
• 负责人: TSUJI Kouichiro
• 金额: $ 9.28万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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/en/grant/KAKENHI-PROJECT-13470157/
• 关键词: Regenerative medicine; Organogenesis; Generation of hematopoiesis; Yolk sac; AGM region; CD34; Hematopoietic stem cell; Fetal therapy; 造血; 全胚胎児培養; 血管内皮細胞; 二次造血; 全胚培養; B細胞; OP9ストローマ細胞; キメラ胎仔マウス

In this project, we indicated the following results according mouse embryonic hematopoiesis.(1)In mouse embryogenesis, the primitive hematopoiesis in 7.5 days postcoitum (dpc) yolk sac (YS) has megakryocytopoiesis different from that of the definitive hematopoiesis in adult bone marrow.(2)Stromal cells derived from aorta-gonad-mesonephros (AGM) region in 10.5 dpc mouse embryo can support the development of hematopoietic stem cells (HSC) in the definitive hematopoiesis.(3)The Stromal cells from AGM region in 10.5 dpc mouse embryo can stimulate the generation of definitive hematopoiesis from not only AGM region but also YS at 8.0 dpc, a time when YS and embryo are not connected by blood vessels.(4)8.25 dpc mouse YS was engrafted upon YS of donor embryo at the same stage to make YS-YS chimeric embryo. After the incubation of the chimeric embryo for 66 hours, we identified B cells originated from donor YS in the chimera, suggesting that a part of definitive hematopoiesis at least is generated from YS in mouse embryogenesis.(5) The stromal cells are also capable of supporting the development of cord blood HSC derived from human fetal hematopopiesis.(6) While CD34 is expressed on c-Kit+ HSC in mouse fetuses and neonates, the expression on HSC decreased with aging, and in mice older than 10 weeks, most of HSC are included in CD34-negative fraction.These results are useful for the further development of regenerative medicine, and the analysis of mammalian organogenesis.

在本项目中，我们根据小鼠胚胎造血得出了以下结果：(1)在小鼠胚胎发生中，交配后7.5天(dpc)卵黄囊(YS)的原始造血具有与成年骨髓中的确定性造血不同的巨冷冻细胞生成。 (2)10.5中源自主动脉-性腺-中肾(AGM)区域的基质细胞dpc小鼠胚胎可以支持定型造血中造血干细胞（HSC）的发育。（3）10.5 dpc小鼠胚胎中来自AGM区的基质细胞不仅可以刺激AGM区而且还可以刺激8.0 YS的定型造血的产生dpc，YS和胚胎没有血管连接的时间。(4)8.25 dpc小鼠YS被移植到YS上供体胚胎在同一阶段制备YS-YS嵌合胚胎。嵌合胚胎孵育66小时后，我们鉴定出嵌合体中源自供体YS的B细胞，这表明在小鼠胚胎发生过程中，至少有一部分确定性造血是由YS产生的。(5)基质细胞还能够支持源自人类胎儿造血的脐带血 HSC 的发育。(6) 虽然 CD34 在小鼠胎儿和新生儿的 c-Kit+ HSC 上表达，但 HSC 上的表达随着衰老，并且在10周以上的小鼠中，大部分HSC包含在CD34阴性部分中。这些结果对于再生医学的进一步发展以及哺乳动物器官发生的分析是有用的。

项目成果

Yoshimasu T., et al.: "Prompt and durable hematopoietic reconstitution by an unrelated cord blood transplantation in a child with Fanconi anemia."Bone Marrow transplant.. 27. 767-769 (2001)

Yoshimasu T. 等人：“通过无关的脐带血移植对患有范可尼贫血的儿童进行迅速和持久的造血重建。”骨髓移植.. 27. 767-769 (2001)

辻浩一郎: "血液細胞の再生医学"ゲノム医学. (印刷中).

Koichiro Tsuji：“血细胞再生医学”基因组医学（正在出版）。

Miyamoto K: "Inhibitory effect of interleukin-3 on the early development of human B-lymphopoiesis"Br J Haematol. 114. 690-697 (2001)

Miyamoto K：“白细胞介素 3 对人类 B 淋巴细胞早期发育的抑制作用”Br J Haematol。

辻浩一郎: "幹細胞を利用した再生医療"小児内科. 34. 37-41 (2001)

Koichiro Tsuji：“使用干细胞的再生医学”小儿内科医学34。37-41（2001）。

Ito M., et al.: "NOD/SCID/gcnull mouse : A novel excellent recipient mouse for engraftment of human cells."Blood. 100. 3175-3182 (2002)

Ito M. 等人：“NOD/SCID/gcnull 小鼠：一种用于植入人类细胞的新型优秀受体小鼠。”血液。