Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies
Abstract Background Endothelial cells (ECs) play a key role in tissue homeostasis, in several pathological conditions, and specifically in the control of vascular functions. ECs are frequently used as in vitro model systems for cardiovascular studies and vascular biology. The porcine model is common...
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| Format: | Article |
| Language: | English |
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BMC
2019-08-01
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| Series: | BMC Molecular and Cell Biology |
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| Online Access: | http://link.springer.com/article/10.1186/s12860-019-0215-2 |
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| author | Olli-Pekka A. Hätinen Johanna E. Lähteenvuo Henna J. Korpela Juho J. Pajula Seppo Ylä-Herttuala |
| author_facet | Olli-Pekka A. Hätinen Johanna E. Lähteenvuo Henna J. Korpela Juho J. Pajula Seppo Ylä-Herttuala |
| author_sort | Olli-Pekka A. Hätinen |
| collection | DOAJ |
| description | Abstract Background Endothelial cells (ECs) play a key role in tissue homeostasis, in several pathological conditions, and specifically in the control of vascular functions. ECs are frequently used as in vitro model systems for cardiovascular studies and vascular biology. The porcine model is commonly used in human clinical cardiovascular studies. Currently, however, there is no robust protocol for the isolation of porcine heart ECs. We have developed a fast isolation protocol, which is cost effective, takes only 1–2 h, and produces EC purity of over 97%. This protocol is optimized for porcine hearts but can be adapted for use with other large animals. Methods Heart is washed by flushing with PBS, whereafter endothelial cells are detached by collagenase incubation and the cells can then be collected immediately after the incubation and plated within an hour after the heart is isolated from a pig. Results The swiftness of the protocol limits changes in the phenotype and RNA expression profile of the cells. Cells were identified as ECs with CD31 (PECAM-1) antibody immunostaining. Functionality of ECs were ensured with in vitro angiogenesis assay. The purity of the ECs was verified by using fluorescence assisted cell sorting (FACS) with the CD31 antibody. Conclusion We developed a new, fast, and cost-effective isolation method for pig heart ECs. Successful isolation of pure ECs is a prerequisite for several cardiovascular and vascular biology studies. |
| first_indexed | 2024-12-23T04:51:33Z |
| format | Article |
| id | doaj.art-420b387bc0384ba88072d0af011404e4 |
| institution | Directory Open Access Journal |
| issn | 2661-8850 |
| language | English |
| last_indexed | 2024-12-23T04:51:33Z |
| publishDate | 2019-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Molecular and Cell Biology |
| spelling | doaj.art-420b387bc0384ba88072d0af011404e42022-12-21T17:59:26ZengBMCBMC Molecular and Cell Biology2661-88502019-08-012011510.1186/s12860-019-0215-2Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studiesOlli-Pekka A. Hätinen0Johanna E. Lähteenvuo1Henna J. Korpela2Juho J. Pajula3Seppo Ylä-Herttuala4A.I. Virtanen Institute for molecular sciences, University of Eastern FinlandA.I. Virtanen Institute for molecular sciences, University of Eastern FinlandA.I. Virtanen Institute for molecular sciences, University of Eastern FinlandA.I. Virtanen Institute for molecular sciences, University of Eastern FinlandA.I. Virtanen Institute for molecular sciences, University of Eastern FinlandAbstract Background Endothelial cells (ECs) play a key role in tissue homeostasis, in several pathological conditions, and specifically in the control of vascular functions. ECs are frequently used as in vitro model systems for cardiovascular studies and vascular biology. The porcine model is commonly used in human clinical cardiovascular studies. Currently, however, there is no robust protocol for the isolation of porcine heart ECs. We have developed a fast isolation protocol, which is cost effective, takes only 1–2 h, and produces EC purity of over 97%. This protocol is optimized for porcine hearts but can be adapted for use with other large animals. Methods Heart is washed by flushing with PBS, whereafter endothelial cells are detached by collagenase incubation and the cells can then be collected immediately after the incubation and plated within an hour after the heart is isolated from a pig. Results The swiftness of the protocol limits changes in the phenotype and RNA expression profile of the cells. Cells were identified as ECs with CD31 (PECAM-1) antibody immunostaining. Functionality of ECs were ensured with in vitro angiogenesis assay. The purity of the ECs was verified by using fluorescence assisted cell sorting (FACS) with the CD31 antibody. Conclusion We developed a new, fast, and cost-effective isolation method for pig heart ECs. Successful isolation of pure ECs is a prerequisite for several cardiovascular and vascular biology studies.http://link.springer.com/article/10.1186/s12860-019-0215-2Porcine heartEndothelial cellIsolationFresh tissueMyocardialPig |
| spellingShingle | Olli-Pekka A. Hätinen Johanna E. Lähteenvuo Henna J. Korpela Juho J. Pajula Seppo Ylä-Herttuala Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies BMC Molecular and Cell Biology Porcine heart Endothelial cell Isolation Fresh tissue Myocardial Pig |
| title | Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies |
| title_full | Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies |
| title_fullStr | Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies |
| title_full_unstemmed | Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies |
| title_short | Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies |
| title_sort | isolation of fresh endothelial cells from porcine heart for cardiovascular studies a new fast protocol suitable for genomic transcriptomic and cell biology studies |
| topic | Porcine heart Endothelial cell Isolation Fresh tissue Myocardial Pig |
| url | http://link.springer.com/article/10.1186/s12860-019-0215-2 |
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