Purification, expansion, and reconstitution of mesodermal stemcells from human bone marrow

人骨髓中胚层干细胞的纯化、扩增和重建

• 批准号: 15591111
• 负责人: KOBAYASHI Masao
• 金额: $ 2.3万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591111/
• 关键词: Mesenchymal Stem Cell; Bone Marrow; Hematopoiesis; ABC-G2; FACS; Mesenchymal Stem Cell; Bone Marrow; 免疫不全マウス; ABC-G2; 異種間移植

Mesenchymal stem Cells were purified from human bone marrow cells based on the expression of CD34,CD38,CD45, anti-glycophorin A, and ABCG2 antibodies by FACS vantage. The frequency of CD45-negative and ABCG2-positive cells in bone marrow was less than 0.01%. The purified CD45-negative and ABCG2-positive cells were cultured under low serum and low glucose medium with appropriate growth factors resulting in the differentiation into bone, chondrocyte, and adipocytes. Furthermore, bone marrow cells were transplanted into immunodeficient mice (NOD/SCID/b2-microglobulin^<null>) to confirm the reconstituting ability in different organs. The cells negative for ABCG2 were reconstiuted as hematopoietic cells differentiating into leukocytes, whereas those positive for ABCG2 migrated into mesodermal tissues. These observations suggest that the expression of ABCG2 may distinguish the origin between hematopoietic stem cells and mesenchymal stem cells.

通过FACS Vantage检测CD34、CD38、CD45、抗血糖蛋白A和ABCG2抗体的表达，从人骨髓细胞中分离纯化骨髓间充质干细胞。骨髓中CD45阴性细胞和ABCG2阳性细胞的比例均小于0.01%。纯化的CD45阴性和ABCG2阳性细胞在低血清和低糖条件下培养，并加入适当的生长因子，诱导分化为骨、软骨和脂肪细胞。此外，将骨髓细胞移植到免疫缺陷小鼠(NOD/SCID/b2-微球蛋白)中，以证实其在不同器官中的重建能力。ABCG2阴性的细胞被重建为向白细胞分化的造血细胞，而ABCG2阳性的细胞迁移到中胚层组织。这些观察结果提示，ABCG2的表达可能区分了造血干细胞和间充质干细胞的来源。

项目成果

小林正夫, 川口浩史: "好中球減少症"小児内科(小児疾患診療のための病態整理2). 35. 1287-1292 (2003)

小林正男、川口宏：《中性粒细胞减少症》儿科内科（儿科疾病治疗病理分类2）35。1287-1292（2003）。

G-CSF製剤,Epo製剤,M-CSF製剤

G-CSF制剂、Epo制剂、M-CSF制剂

• 发表时间: 2004
• 期刊: 小児科診療 67
• 作者: 小林正夫;佐藤 貴;上田一博
• 通讯作者: 上田一博

Neutropenia in childhood.

儿童时期出现中性粒细胞减少症。

• 发表时间: 2005
• 期刊: Japanese Journal of the Japan Pediatric Society 109
• 作者: 川口浩史;中村和洋;小林正夫;Zilfalil BA;Kobayashi M
• 通讯作者: Kobayashi M

Significance of human neutrophil antigen-2a (NB1) expression and neutrophil number in pregnancy.

人类中性粒细胞抗原 2a (NB1) 表达和中性粒细胞数量在妊娠中的意义。

• 发表时间: 2004
• 期刊: Transfusion 44
• 作者: Taniguchi K;Kobayashi M;et al.
• 通讯作者: et al.

Kobayashi M, Sato T, et al.: "Efficacy of the prophylactic use of trimethoprim-sulfamethoxazole in auotimmune neutropenia in infancy"Journal of Pediatric Hematology/Oncology. 25. 553-557 (2003)

Kobayashi M、Sato T 等人：“预防性使用甲氧苄啶-磺胺甲恶唑治疗婴儿期自身免疫性中性粒细胞减少症的功效”儿科血液学/肿瘤学杂志。