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首页 > 过刊浏览>2020年第30卷第2期 >54-58. DOI:10. 3969 / j.issn.1671-7856. 2020. 02. 008
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GFP 标记脐带间充质干细胞治疗 mdx 小鼠的药代动力学研究
作者:
  • 庞荣清 1

    庞荣清

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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  • 朱 慧

    朱 慧

    1.联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032; 2.延安大学医学院,陕西 延安 716000
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  • 鄢东海

    鄢东海

    1.联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032; 3.巴东县人民医院,湖北 巴东 444300
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  • 王 强 1

    王 强

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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  • 李自安 1

    李自安

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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  • 王金祥 1

    王金祥

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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  • 于倩倩 1

    于倩倩

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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  • 阮光萍 1

    阮光萍

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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  • 朱向情 1

    朱向情

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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  • 潘兴华 1

    潘兴华

    联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032
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作者单位:

1.联勤保障部队第九二〇医院细胞生物治疗中心,干细胞与免疫细胞生物医药技术国家地方联合工程实验室, 云南省细胞治疗技术转化医学重点实验室,云南省干细胞工程实验室,昆明 650032; 2.延安大学医学院,陕西 延安 716000; 3.巴东县人民医院,湖北 巴东 444300

中图分类号:

R-33


Pharmacokinetics study of GFP-labeled mouse umbilical cord mesenchymal stem cells in mdx mice
Author:
  • PANG Rongqing 1

    PANG Rongqing

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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  • ZHU hui

    ZHU hui

    1.Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032,China. 2. Medical school of Yanan universit, Yanan 716000
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  • YAN Donghai

    YAN Donghai

    1.Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032,China. 3. Badong People hospital, Badong 444300
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  • WANG Qiang 1

    WANG Qiang

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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  • LI Zian 1

    LI Zian

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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  • WANG Jinxiang 1

    WANG Jinxiang

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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  • YU Qianqian 1

    YU Qianqian

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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  • RUAN Guangping 1

    RUAN Guangping

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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  • ZHU Xiangqing 1

    ZHU Xiangqing

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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  • PAN Xinghua 1

    PAN Xinghua

    Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032, China
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Affiliation:

1.Cell Biological Therapy Center of 920th Hospital of Joint Logistics Support Force, Integrated Engineering laboratory of Cell Biological Medicine of State and Regions, Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunan Province, Stem Cell Engineering laboratory of Yunnan Province, Kunming 650032,China. 2. Medical school of Yanan universit, Yanan 716000. 3. Badong People hospital, Badong 444300

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    摘要:

    目的 应用小鼠脐带间充质干细胞(mUCMSC)示踪方法和 mdx 小鼠研究细胞药物的药代动力学。 方法 GFP 转染标记 mUCMSC,局部注射 0. 4 mL(5×106 个 GFP-mUCMSC)细胞悬液到腓肠肌和膈肌损伤的 mdx 小鼠腹腔,在 1 h、3 h、5 h、24 h、1 周等 5个时间点用活体成像系统观察小鼠体内的荧光信号,观察结束后处死小 鼠,采集膈肌制作冰冻切片并在荧光显微镜下观察荧光信号。 结果 mUCMSC 与 MOI 为 150 的病毒载体共培养 72 h 可以成功标记 GFP。 局部腹腔注射 0. 4 mL(5×106 个 GFP-mUCMSC)后 1 h,用活体成像系统观察到 mdx 小鼠 腹壁下注射细胞的部位有明亮的绿色荧光集中分布,随着时间延长,荧光面积逐渐减小,24 h 后荧光强度明显变 弱,荧光面积大幅缩小,一周后荧光信号几乎消失;此时,在荧光显微镜下仍可观察到 mdx 小鼠膈肌内存在荧光信 号,与未注射细胞的对照组有显著差异(P﹤0. 05)。 结论 注射到腹腔的脐带间充质干细胞可以进入损伤的膈 肌,其 GFP 荧光信号 1 周内逐渐消失,活体成像系统是监测干细胞体内动态分布和清除率的有效工具。

    Abstract:

    Objective The pharmacokinetics of cellular drugs were studied with the mouse umbilical cord mesenchymal stem cells ( mUCMSC) tracing method and mdx mice. Methods mUCMSC were labeled by GFP transfection, and a 0. 4 mL (5×106 GFP-mUCMSC) cell suspension was injected locally into the abdominal cavity of mdx mice with damaged gastrocnemius and diaphragma. The fluorescence signal in the mice was observed with a live imaging system at five timepoints including 1 h, 3 h, 5 h, 24 h, and 1 wk post injection. The mice were euthanized and their diaphragms were collected to obtain frozen sections for fluorescence microscopy. Results mUCMSCs were successfully labeled when they were co-cultured with the viral vector at an MOI of 150 for 72 h. One hour after local intraperitoneal injection of the GFP-mUCMSCs, a strong green fluorescent signal was observed with the live imaging system under the injection position on the abdominal wall of the mdx mice, and the fluorescent area gradually decreased with time. After 24 h, the fluorescence intensity was obviously weakened, and the fluorescent area was greatly reduced. These fluorescent signals had almost disappeared after one week. At this time, the fluorescent signals could still be observed in the diaphragm of the mdx mice under the fluorescence microscope, and this signal was significantly higher than that of the control group without cell injection (P<0.05). Conclusions Umbilical cord mesenchymal stem cells injected into the abdominal cavity of the mdx mice entered the injured diaphragm and their GFP fluorescent signals gradually disappeared within 1 week. Thus, a live imaging system is an effective tool for monitoring the dynamic distribution and clearance rate of stem cells in vivo.

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庞荣清,朱 慧,鄢东海,王 强,李自安,王金祥,于倩倩,阮光萍,朱向情,潘兴华. GFP 标记脐带间充质干细胞治疗 mdx 小鼠的药代动力学研究[J].中国比较医学杂志,2020,30(2):54~58.

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  • 收稿日期:2019-04-26
  • 在线发布日期: 2020-04-01
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