Study of newly autologous chondrocyte implantation using by gene transfer of hyaluronan synthase

透明质酸合酶基因转移新自体软骨细胞移植的研究

• 批准号: 13671491
• 负责人: WADA Yuichi
• 金额: $ 2.18万
• 依托单位: Chiba 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-13671491/
• 关键词: hyaluronan; gene transfer; chondrocyte implantation; ATDC5; retrovirus vector; ヒアルロン酸合成酵素

In the present study, plasmids containg HAS2 under the PMX-IRES-GFP retrovirus vector were transfected into undifferentiated ATDC5. As a control, cells transfected with the empty vector were used. ATDC5 cells differentiated in the presence of insulin and RT-PCR was performed to detect HAS2 m RNA expression and we evaluated visualization of pericellular coats which were composed of newly synthesized HA using by immunofluorescent staining and analyzed HA concentration in culture medium by high performance liquid chromatography. The signal of the cells transfected with HAS2 was visually much stronger than that of control cells. The HA concentration of the cells transfected with HAS2 in culture medium was much higher than that of control throughout 21 days. By transfection of HAS2 with retrovirus, we are able to expect the continuous synthesis of HA. Therefore this technique may be a great significance in autologous chondrocyte implantation.

在本研究中，将PMX-IRES-GFP逆转录病毒载体下的HAS2抑制质粒转染为未分化的ATDC5。作为对照，使用了用空载体转染的细胞。在存在胰岛素和RT-PCR的情况下进行分化的ATDC5细胞检测HAS2 M RNA表达，我们评估了细胞细胞层的可视化，这些层由高性能液体液体分析，由免疫荧光染色和培养基中分析的HA浓度在培养基中分析的HA浓度。用HAS2转染的细胞的信号在视觉上比对照细胞的信号强得多。在培养基中，用HAS2转染的细胞的HA浓度在21天内远高于对照的HA浓度。通过用逆转录病毒转染HAS2，我们能够期望HA的连续合成。因此，该技术在自体软骨细胞植入中可能具有重要意义。

项目成果

K.Tsuchiya, Y.Wada, K.Takase, K.Nakagawa, T.Yamashita, A.Watanabe and H.Moriya: "EVALUATION OF HYALURONAN SYNTHESIS AND EXPRESSION OF CD44 BY GENE TRANSFER OF HYALURONAN SYNTHASE 2"Osteoarthritis and Cartilage. 10 supplement B. (2002)

K.Tsuchiya、Y.Wada、K.Takase、K.Nakakawa、T.Yamashita、A.Watanabe 和 H.Moriya：“通过透明质酸合成酶 2 的基因转移评估透明质酸合成和 CD44 的表达”骨关节炎和软骨。

Kan Tsuchiya, Yuichi Wada, et al.: "EVALUATION of HYALURONAN SYNTHESIS BY RETROVIRUS-MEDIATED GENE TRANSFER of HYALURONAN"Osteoarthritis and Cartilage. Vol.10. SB (2002)

Kan Tsuchiya、Yuichi Wada 等人：“通过逆转录病毒介导的透明质酸基因转移来评估透明质酸合成”骨关节炎和软骨。

Kan Tsuchiya ,Yuichi Wada, Kan Takase, Kouichi Nakagawa, Takeshi Yamashita and Hideshige Moriya: "EVALUATION OF HYALURONAN SYNTHESIS BY RETROVIRUS-MEDIATED GENE TRANSFER OF HYALURONAN 2"J. Jpn. Orthop. Assoc.. 76(8). S1095 (2002)

Kan Tsuchiya、Yuichi Wada、Kan Takase、Kouichi Nakakawa、Takeshi Yamashita 和 Hideshige Moriya：“通过逆转录病毒介导的透明质酸 2 基因转移来评估透明质酸合成”J。

Kan Tsuchiya, Yuichi Wada, Kan Takase, Kouichi Nakagawa, Takeshi Yamashita, Hiroki Sakai, Norimasa Takahashi, Atsuya Watanabe, Rie Koh, Yoshikuni Kawaguchi and Hideshige Moriya: "EVALUATION OF HYALURONAN SYNTHESIS BY RETROVIRUS-MEDIATED GENE TRANSFER OF H

Kan Tsuchiya、Yuichi Wada、Kan Takase、Kouichi Nakakawa、Takeshi Yamashita、Hiroki Sakai、Norimasa Takahashi、Atsuya Watanabe、Rie Koh、Yoshikuni Kawaguchi 和 Hideshige Moriya：“通过逆转录病毒介导的 H 基因转移来评估透明质酸合成”

Kan Tsuchiya, Yuichi Wada, et al.: "Evaluation of Hyaluronan Synthesis by Retrovirus-Mediated Gene Transfer of Hyaluronan"Osteoarthritis and Cartilage. Vol.10. S.B. (2002)

Kan Tsuchiya、Yuichi Wada 等人：“通过逆转录病毒介导的透明质酸基因转移来评估透明质酸合成”骨关节炎和软骨。