Isolation and Purification of Murine Cardiac Pericytes
| Autor: | Vishnu Chintalgattu, Linda L. Lee, Aarif Y. Khakoo |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Angiogenesis General Chemical Engineering Cell Cell Culture Techniques Cell Separation 030204 cardiovascular system & hematology General Biochemistry Genetics and Molecular Biology Mice 03 medical and health sciences 0302 clinical medicine Antigen Antigens CD medicine Animals Primary cell General Immunology and Microbiology Chemistry Myocardium General Neuroscience Cell sorting Cardiovascular physiology Cell biology 030104 developmental biology medicine.anatomical_structure Cell culture CD146 Pericytes Biomarkers |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| Popis: | Pericytes, perivascular cells of microvessels and capillaries, are known to play a part in angiogenesis, vessel stabilization, and endothelial barrier integrity. However, their tissue-specific functions in the heart are not well understood. Moreover, there is currently no protocol utilizing readily accessible materials to isolate and purify pericytes of cardiac origin. Our protocol focuses on using the widely used mammalian model, the mouse, as our source of cells. Using the enzymatic digestion and mechanical dissociation of heart tissue, we obtained a crude cell mixture that was further purified by fluorescence activating cell sorting (FACS) by a plethora of markers. Because there is no single unequivocal marker for pericytes, we gated for cells that were CD31-CD34-CD45-CD140b+NG2+CD146+. Following purification, these primary cells were cultured and passaged multiple times without any changes in morphology and marker expression. With the ability to regularly obtain primary murine cardiac pericytes using our protocol, we hope to further understand the role of pericytes in cardiovascular physiology and their therapeutic potential. |
| Databáze: | OpenAIRE |
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