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中华肾病研究电子杂志

中华肾病研究电子杂志 ›› 2021, Vol. 10 ›› Issue (06) : 323 -327. doi: 10.3877/cma.j.issn.2095-3216.2021.06.005

论著

小鼠肾脏包膜下移植间充质干细胞方法探索
李铎1, 黄梦杰2, 陈尚3, 李宗金3, 陈健文2,†(), 陈香美4,†()   
  1. 1. 100853　北京，解放军总医院第一医学中心肾脏病科、解放军肾脏病研究所、肾脏疾病国家重点实验室、国家慢性肾病临床医学研究中心、肾脏疾病研究北京市重点实验室；300071　天津大学应急医学研究院；453003　新乡医学院第一附属医院肿瘤科
    2. 100853　北京，解放军总医院第一医学中心肾脏病科、解放军肾脏病研究所、肾脏疾病国家重点实验室、国家慢性肾病临床医学研究中心、肾脏疾病研究北京市重点实验室
    3. 300071　天津，南开大学医学院
    4. 100853　北京，解放军总医院第一医学中心肾脏病科、解放军肾脏病研究所、肾脏疾病国家重点实验室、国家慢性肾病临床医学研究中心、肾脏疾病研究北京市重点实验室；300071　天津，南开大学医学院
  • 收稿日期:2021-08-16 出版日期:2021-12-28
  • 通信作者: 陈健文, 陈香美
  • 基金资助:
    国家自然科学基金重点项目(82030025); 国家自然科学基金青年项目(82000631、82100713); 中国博士后科学基金特别资助(2021T140791)

Exploration of the method to transplant mesenchymal stem cells under the capsule of mouse kidney

Duo Li1, Mengjie Huang2, Shang Chen3, Zongjin Li3, Jianwen Chen2,†(), Xiangmei Chen4,†()   

  1. 1. Department of Nephrology, First Medical Center of Chinese PLA General Hospital, Chinese PLA Institute of Nephrology, State Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Diseases, Beijing 100853; Tianjin University Institute of Disaster and Emergency Medicine, Tianjin 300071; Department of Oncology, First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province
    2. Department of Nephrology, First Medical Center of Chinese PLA General Hospital, Chinese PLA Institute of Nephrology, State Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Diseases, Beijing 100853
    3. Nankai University School of Medicine, Tianjin 300071; China
    4. Department of Nephrology, First Medical Center of Chinese PLA General Hospital, Chinese PLA Institute of Nephrology, State Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Diseases, Beijing 100853; Nankai University School of Medicine, Tianjin 300071; China
  • Received:2021-08-16 Published:2021-12-28
  • Corresponding author: Jianwen Chen, Xiangmei Chen
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引用本文：

李铎, 黄梦杰, 陈尚, 李宗金, 陈健文, 陈香美. 小鼠肾脏包膜下移植间充质干细胞方法探索[J]. 中华肾病研究电子杂志, 2021, 10(06): 323-327.

Duo Li, Mengjie Huang, Shang Chen, Zongjin Li, Jianwen Chen, Xiangmei Chen. Exploration of the method to transplant mesenchymal stem cells under the capsule of mouse kidney[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2021, 10(06): 323-327.

目的

探索肾脏包膜下移植间充质干细胞(MSCs)，以期提高细胞有效移植量和留滞率。

方法

首先用墨水对小鼠肾脏分别进行实质注射和包膜下注射初步探究其效果和可行性；然后将MSCs与胶原基质共培养以探究细胞存活和生长情况。将实验小鼠分为胶原/MSCs的肾实质移植组和肾包膜下移植组，对移植后的肾脏进行称重、HE染色观察。肾包膜下移植小鼠又分为单一胶原移植组和胶原与MSCs混合移植组，在术后不同时间点进行活体成像观察。

结果

与肾实质注射相比，肾包膜下注射的墨水可以覆盖更大范围的肾组织，并且有更大的移植容量。MSCs在胶原基质中生长状态良好；肾包膜下注射胶原与MSCs混悬液后，MSCs呈现出有效、稳定的移植量，并且留滞更长时间。

结论

肾包膜下移植表明，胶原包裹的MSCs移植量稳定、且可长时间留滞、有一定可行性。此研究为MSCs移植治疗肾损伤提供了新思路，具有潜在的临床应用价值。

关键词: 小鼠, 干细胞移植, 肾包膜, 留滞率
Objective

To explore the transplantation of mesenchymal stem cells (MSCs) under the kidney capsule for the purpose of increasing the effective transplantation volume and retention rate of cells.

Methods

Firstly, parenchymal injection and subcapsular injection of mouse kidneys were performed with ink respectively to preliminarily explore the effect and feasibility. Then the MSCs were co-cultured with collagen matrix to investigate the conditions of cell survival and growth. The collagen/MSCs transplantation mice were divided into renal parenchymal transplantation group and renal subcapsular transplantation group. After transplantation, the kidneys were weighed and observed with HE staining. Mice of the renal subcapsular transplantation group were again divided into single collagen transplantation group and collagen plus MSCs transplantation group. In vivo imaging observations were performed at different time points after the operation.

Results

Compared with renal parenchymal injection, renal subcapsular injection made the ink cover a larger range of renal tissue and had a bigger bearing capacity. The MSCs grew well in the collagen matrix. After the renal subcapsular injection of collagen and MSCs suspension, the MSCs showed an effective and stable transplantation amount, and remained for a longer time.

Conclusion

The renal subcapsular transplantation showed that the amount of collagen-wrapped MSCs was stable and retained for a longer time with a certain feasibility. This study provided a new idea for MSCs transplantation in treating kidney injury, being of potential value for clinical application.

Key words: Mice, Stem cell transplantation, Renal capsule, Retention rate
图1 小鼠肾脏墨水注射对比注：A：空白对照组；B：肾实质注射组；C：肾包膜下注射组
图2 胶原性状、结构和胶原/MSCs共培养注：A：玻璃小瓶盛装不同温度下的胶原形态；B：胶原结构电镜照片(×2 000)；C：胶原/MSCs共聚焦小皿共培养；D：MSCs正常贴壁培养；E：胶原/MSCs共培养0 h；F：胶原/MSCs共培养48 h
图3 胶原/ MSCs实质、包膜下移植对比注：A：肾脏增加质量比较；B：肾脏质量增加百分比比较；C：未进行包膜下注射包膜状态PAS染色显示；D：包膜下注射后包膜状态PAS染色显示；与实质注射组比较，a P<0.05
图4 细胞留滞情况观察注：A：梯度种植标记Fluc的MSCs；B：细胞数与Fluc表达的相关性；C：小动物活体成像显示标记细胞在14 d内的示踪；D：随时间变化的Fluc信号强度统计量化
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