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PriCells: Isolation, culture, and purification of Human Pulmonary Arterial Smooth Muscle Cells
PriCells: Isolation, culture, and purification of Human Pulmonary Arterial Smooth Muscle Cells (HPASMCs)
1. HPASMCs were isolated from lungs resected from tissue of normal human or adult patients with malignant tumors.
2. The arteries were separated from their adventitia and endothelium and then minced into 1- to 3-mm2 pieces with sterile scalpel blades.
3. The tissues were mounted in tissue culture wells.
4. The HPASMCs pellets were resuspended in Pricells Culture System and c*ted on 6-well plates pre-coated with gelatin (0.2%) for several days until subconfluent.
5. Purification: The HPASMCs were isolated using magnetic beads (Dynal) according to the instructions of the manufacturer.
6. The medium was replaced every 2 to 3 days, and the subcultures were obtained by trypsin/EDTA treatment of confluent monolayers at a splitting ratio of 1:3.
7. Identification: Cells were identified with PriCells Identification System.