Hello Sara Abd Al Rahim

Fibroblasts attach to the substratum much quickly (say within 2 hours), while the smooth muscle cells may remain floating in the medium. So, you may aspirate the supernatant which will contain your cells of interest and may also contain a few fibroblasts. You may further purify your primary cells by repeated plating of cells and performing the same steps as above until no further fibroblasts are detected in the pulmonary artery smooth muscle culture. You may confirm the purity of your primary culture by positive staining for specific markers.

Alternatively, fibroblasts detach very quickly on mild trypsinization. So, you could use mild trypsin treatment and get rid of fibroblasts, while the primary pulmonary artery smooth muscle cells would still remain attach to the substratum.

Still another way out would be the use of magnetic cell sorting which is a well-established method for the separation of various cell populations based on specific cell surface antigens and therefore allows for the enrichment of a particular cell population. In principle, specific antibodies bound to magnetic beads capture cells which express the corresponding epitope. Using a magnetic field, the labeled cells are retained and therefore separated from unlabeled cells. Cells may be isolated via negative selection (labeling of undesired cells) or positive selection (labeling of target cells), depending on the particular antigen that is used.

Best.

Thank you Malcolm Nobre