A modified primary culture method of rat pulmonary vein smooth muscle cells
Abstract Background Although the pressure of pulmonary vein increases before pulmonary artery in pulmonary hypertension due to left heart disease (PH-LHD), only a few studies have assessed pulmonary vein smooth muscle cells (PVSMCs) because of the lack of a simple and feasible isolation method. Meth...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-04-01
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| Series: | Journal of Cardiothoracic Surgery |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13019-023-02233-1 |
| _version_ | 1797789550268907520 |
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| author | Wenhui Huang Hongjin Liu Yichao Pan Xueying Wang Hongwei Yang Danjie Wang Jing Lin Hui Zhang |
| author_facet | Wenhui Huang Hongjin Liu Yichao Pan Xueying Wang Hongwei Yang Danjie Wang Jing Lin Hui Zhang |
| author_sort | Wenhui Huang |
| collection | DOAJ |
| description | Abstract Background Although the pressure of pulmonary vein increases before pulmonary artery in pulmonary hypertension due to left heart disease (PH-LHD), only a few studies have assessed pulmonary vein smooth muscle cells (PVSMCs) because of the lack of a simple and feasible isolation method. Methods In this study, we introduced a simple method to obtain PVSMCs. Primary pulmonary veins were removed by puncture needle cannula guidance. Then, PVSMCs were cultured by the tissue explant method and purified by the differential adhesion method. The cells were characterized by hematoxylin-eosin (HE) staining, immunohistochemistry, western blotting, and immunofluorescence to observe the morphology and verify the expression of alpha-smooth muscle actin (α-SMA). Results The HE staining results showed that the pulmonary vein media was thinner than the pulmonary artery, the intima and adventitia of the pulmonary vein were removed by this method, and the obtained cells with good activity exhibited morphological characteristics of smooth muscle cells. In addition, higher α-SMA expression was observed in the cells obtained by our isolation method than in the traditional method. Conclusion This study established a simple and feasible method to isolate and culture PVSMCs that might facilitate the cytological experiments for PH-LHD. |
| first_indexed | 2024-03-13T01:52:16Z |
| format | Article |
| id | doaj.art-2e5de101ba7749b481cd83ee74aceae3 |
| institution | Directory Open Access Journal |
| issn | 1749-8090 |
| language | English |
| last_indexed | 2024-03-13T01:52:16Z |
| publishDate | 2023-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Cardiothoracic Surgery |
| spelling | doaj.art-2e5de101ba7749b481cd83ee74aceae32023-07-02T11:26:38ZengBMCJournal of Cardiothoracic Surgery1749-80902023-04-011811810.1186/s13019-023-02233-1A modified primary culture method of rat pulmonary vein smooth muscle cellsWenhui Huang0Hongjin Liu1Yichao Pan2Xueying Wang3Hongwei Yang4Danjie Wang5Jing Lin6Hui Zhang7Critical Care Medicine, Union Hospital of Fujian Medical UniversityDepartment of Cardiovascular Surgery, Union Hospital of Fujian Medical UniversityDepartment of Cardiovascular Surgery, Union Hospital of Fujian Medical UniversityCritical Care Medicine, Union Hospital of Fujian Medical UniversityDepartment of Cardiovascular Surgery, Union Hospital of Fujian Medical UniversityCritical Care Medicine, Union Hospital of Fujian Medical UniversityDepartment of Cardiovascular Surgery, Union Hospital of Fujian Medical UniversityCritical Care Medicine, Union Hospital of Fujian Medical UniversityAbstract Background Although the pressure of pulmonary vein increases before pulmonary artery in pulmonary hypertension due to left heart disease (PH-LHD), only a few studies have assessed pulmonary vein smooth muscle cells (PVSMCs) because of the lack of a simple and feasible isolation method. Methods In this study, we introduced a simple method to obtain PVSMCs. Primary pulmonary veins were removed by puncture needle cannula guidance. Then, PVSMCs were cultured by the tissue explant method and purified by the differential adhesion method. The cells were characterized by hematoxylin-eosin (HE) staining, immunohistochemistry, western blotting, and immunofluorescence to observe the morphology and verify the expression of alpha-smooth muscle actin (α-SMA). Results The HE staining results showed that the pulmonary vein media was thinner than the pulmonary artery, the intima and adventitia of the pulmonary vein were removed by this method, and the obtained cells with good activity exhibited morphological characteristics of smooth muscle cells. In addition, higher α-SMA expression was observed in the cells obtained by our isolation method than in the traditional method. Conclusion This study established a simple and feasible method to isolate and culture PVSMCs that might facilitate the cytological experiments for PH-LHD.https://doi.org/10.1186/s13019-023-02233-1Pulmonary vein smooth muscle cellsPulmonary veinsPrimary cell culturePulmonary hypertension due to left heart disease |
| spellingShingle | Wenhui Huang Hongjin Liu Yichao Pan Xueying Wang Hongwei Yang Danjie Wang Jing Lin Hui Zhang A modified primary culture method of rat pulmonary vein smooth muscle cells Journal of Cardiothoracic Surgery Pulmonary vein smooth muscle cells Pulmonary veins Primary cell culture Pulmonary hypertension due to left heart disease |
| title | A modified primary culture method of rat pulmonary vein smooth muscle cells |
| title_full | A modified primary culture method of rat pulmonary vein smooth muscle cells |
| title_fullStr | A modified primary culture method of rat pulmonary vein smooth muscle cells |
| title_full_unstemmed | A modified primary culture method of rat pulmonary vein smooth muscle cells |
| title_short | A modified primary culture method of rat pulmonary vein smooth muscle cells |
| title_sort | modified primary culture method of rat pulmonary vein smooth muscle cells |
| topic | Pulmonary vein smooth muscle cells Pulmonary veins Primary cell culture Pulmonary hypertension due to left heart disease |
| url | https://doi.org/10.1186/s13019-023-02233-1 |
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