Detailed Study during Cartilage Formation by Human Mesenchymal Stern Cells In a Three Dimentional Culture and Implantation.

三维培养和植入中人间充质干细胞软骨形成过程的详细研究。

• 批准号: 14580813
• 负责人: ICHINOSE Shizuko
• 金额: $ 2.43万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580813/
• 关键词: Human mesenchymal stem cells; Chondrogenesis induction; Alginate bead culture; Pellet culture; Implantation; Cartilage formation; ヒト骨髄由来間葉系幹細胞; アルジネート培養; TGFβ-3

Human mesenchymal stem cells (hMSCs) have the potential to differentiate into the lineages of mesenchymal tissues. The aim of this study Is to clarify the detailed process of differentiation In hMSCs. We examined cartilage formation of human mesenchymal stem cells using three dimentional culture and implantation.Three dimentional culture -alginate bead culture-: hMSCs were maintained in alginate beads. We examined them by light microscope, transmission electron microscope and immunocryo-ultramicrotomy. hMSCs produced collagen type II, X, proteoglycan and osteocalcin at day 4 in the case of alginate bead culture. Thereafter electron-dense particles, composed of hydroxyapatite, appeared.Three dimentional culture -pellet culture-: In the case of pellet culture, hMSCs were connected with junctional complex at day 1. At day 7, hMSCs produced collagen type I, II, X and proteoglycan. During day 14-21, the expression of collagen type II decreased.Implantation of MSO-derived cartilages into the rabbit cartilage defect.: We cultured MSCs as a micromass in chondrogenesis medium for 14 days and differentiated MSCs into cartilage in vitro. Thereafter, MSC-derived cartilages were press fitted into the rabbit cartilage defects. Eight weeks after implantation, we observed the cartilage defects were repaired.

人骨髓间充质干细胞(HMSCs)具有向间充质组织分化的潜能。本研究旨在阐明人骨髓间充质干细胞分化的详细过程。我们用三维培养和植入的方法检测了人骨髓间充质干细胞的软骨形成。三维培养-海藻酸珠培养-hMSCs在海藻酸珠中维持。光镜、透射电子显微镜和免疫结晶超微结构观察。在藻酸盐珠培养的第4天，hMSCs产生了II型、X型胶原、蛋白多糖和骨钙素。三维培养-颗粒培养：颗粒培养第1天，hMSCs与连接复合体相连；第7天，hMSCs产生I型、II型、X型胶原和蛋白多糖。在第14-21天，II型胶原的表达减少。将MSO来源的软骨移植到兔软骨缺损处。：我们将MSCs作为微团在软骨形成液中培养14天，并在体外将MSCs分化为软骨。然后，将MSC来源的软骨按压植入兔软骨缺损处。植入8周后，观察软骨缺损修复情况。

项目成果

市野瀬志津子, 宗田 大, 他2名: "ヒト骨髄由来間葉系幹細胞を用いた軟骨組織の分化誘導"日本顕微鏡学会第59回学術講演会発表要旨集. Vol.38,Supplement 1. 197 (2003)

Shizuko Ichinose、Dai Muneta 等 2 人：“利用人骨髓来源的间充质干细胞诱导软骨组织的分化”日本显微镜学会第 59 届学术会议摘要，增刊 1. 197（2003 年） ) )

Ichinose S, Muneta T et al.: "TEM observation of seven retrieved total knee Joints made of Co-Cr-Mo and Ti-Ai-v alloys"Blo-Medical Materials and Engineering. val.13. 125-134 (2003)

Ichinose S、Muneta T 等人：“七个回收的 Co-Cr-Mo 和 Ti-Al-V 合金全膝关节的 TEM 观察”Blo-医疗材料与工程。

M.Tagami, S.Ichinose, et al.: "Genetic and ultrastructural demonstaration of strong reversibility in human mesenchymal stem cell"Cell tissue Res 312,31-40 (2003). 31. 31-40 (2003)

M.Tagami、S.Ichinose 等：“人间充质干细胞强可逆性的遗传和超微结构论证”细胞组织 Res 312,31-40 (2003)。

S.ICHINOSE, T.MUNETA, et al.: "The Study of Metal Ion Release and Cytotoxicity in Co-Cr-Mo and Ti-Al-V Alloy in Total Knee Prosthesis-Scanning Electron Microscopic Observation"JOURNAO OF MATERIALS SCIENCE : MATERIALS IN MEDICINE. 14. 79-86 (2003)

S.ICHINOSE、T.MUNETA 等人：“全膝关节假体中 Co-Cr-Mo 和 Ti-Al-V 合金中金属离子释放和细胞毒性的研究 - 扫描电子显微镜观察”材料科学杂志：材料

S.Kawano, S.Shoji, S.Ichinose, et al.: "Characterization of Ca^2 Signaling pathways in human mesenchymal stem cells"Cell Calcium. 32. 165-174 (2002)

S.Kawano、S.Shoji、S.Ichinose 等人：“人间充质干细胞中 Ca^2 信号通路的表征”细胞钙。