Primary Culture of Rat Aortic Vascular Smooth Muscle Cells: A New Method.
Primary Culture of Rat Aortic Vascular Smooth Muscle Cells: A New Method.
复制标题
大鼠主动脉血管平滑肌细胞的原发性培养：一种新方法。
DOI：
10.12659/msm.902816
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发表时间：
2017-08-19
期刊：
影响因子：
--
通讯作者：
Guo H
中科院分区：
文献类型：
--
作者：
Chi J;Meng L;Pan S;Lin H;Zhai X;Liu L;Zhou C;Jiang C;Guo H
开发一种简单有效的获得原代培养VSMCs的方法是基础心血管研究的必要条件。我们的新方法主要包括6个步骤：分离主动脉，去除动脉周围的脂肪组织，分离培养基，将培养基切割成小的组织块，将组织块转移到细胞培养板上，孵育至细胞汇合。细胞形态和免疫荧光鉴定为VSMCs。将该组织外植体法、传统组织外植体法、酶消化法和A7r5细胞系获得的VSMCs分为4组。多次荧光染色检测细胞纯度。采用Western blotting检测不同方法获得的VSMCs的表型。细胞在8天左右开始生长，在16天内变得相对融合。与传统组织外植法、酶切法或A7r5细胞系获得的VSMCs相比，本方法获得的VSMCs纯度更高，且表现出更“收缩”的表型特征。克服了以往原代培养方法的不足，建立了一种简单可靠、纯度高、稳定性好的大鼠主动脉VSMCs分离培养方法。
Developing a simple and efficient method of obtaining primary cultured VSMCs is necessary for basic cardiovascular research. The procedure of our new method mainly includes 6 steps: isolation of the aortic artery, removal of the fat tissue around the artery, separation of the media, cutting the media into small tissue blocks, transferring the tissue blocks to cell culture plates, and incubation until the cells reach confluence. The cells were identified as VSMCs by morphology and immunofluorescence. Then, VSMCs obtained by this new tissue explants method, the traditional tissue explants method, the enzyme digestion method, and A7r5 cell line were divided into 4 groups. The purity of cells was test by multiple fluorescent staining. Western blotting was used to investigate the phenotype of VSMCs obtained by different methods. Cells began to grow out at about 8 days and became relatively confluent within 16 days. Compared with VSMCs from the traditional tissue explants method and enzyme digestion method or A7r5 cell line, VSMCs obtained by our method showed higher purity and manifested a more “contractile” phenotype characteristic. We have conquered the disadvantages in the previous primary culture methods and established a simple and reliable way to isolate and culture rat aortic VSMCs with high purity and stability.
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影响因子：
6.2
作者：
Moussallem, Maroun D.;Olenych, Scott G.;Scott, Shannon L.;Keller, Thomas C. S., III;Schlenoff, Joseph B.
通讯作者：
Schlenoff, Joseph B.
影响因子：
5.8
作者：
Torres G;Morales PE;García-Miguel M;Norambuena-Soto I;Cartes-Saavedra B;Vidal-Peña G;Moncada-Ruff D;Sanhueza-Olivares F;San Martín A;Chiong M
通讯作者：
Chiong M
影响因子：
--
作者：
Halidi, Nadia;Alonso, Florian;Meister, Jean-Jacques
通讯作者：
Meister, Jean-Jacques
影响因子：
39.3
作者：
Peeters, Wouter;Moll, Frans L.;Pasterkamp, Gerard
通讯作者：
Pasterkamp, Gerard
影响因子：
3.7
作者：
Simard E;Söllradl T;Maltais JS;Boucher J;D'Orléans-Juste P;Grandbois M
通讯作者：
Grandbois M