Rat aortic smooth muscle cell culture can be used to (1) obtain rat aortic smooth muscle cells, (2) for vascular disease research, and (3) for arterial disease research.

Operation method

chunking

Principle

SD rat thoracic aortic smooth muscle cell culture was performed using the tissue block apposition method, and the cultured cells were visualized by inverted phase contrast microscopy, observed morphologically after HE staining, and identified by immunohistochemistry.

Materials and Instruments

SD rat
PBS FBS DMEM
Surgical scissors Surgical forceps Ophthalmic scissors Ophthalmic forceps Large scalpel Scalpel Petri dish

Move

I. Experimental steps

1. SD rats (150-200 g), severed heads, were immersed in 75% alcohol for 15 min.
2. Place the rats in the ultra-clean table, fix the rats on the rodenticide plate, open the abdominal cavity under sterile conditions, separate the thoracic aorta and abdominal aorta, and extract them.
3. Wash with PBS for 3 times, peel off the outer membrane, scrape off the inner membrane with a scalpel blade, leave the middle membrane and transfer it into 1 mL of medium, cut it into 1x1 ^{mm2 sized} tissue pieces.
4. Wash the tissue blocks with PBS, transfer them into 25 mL culture flasks, stick the tissue blocks evenly with a sharp-nosed elbow, and place them into an incubator at 37℃ with 5% _CO2 for 6 hours, and then add 2.5 mL of culture medium after the tissue blocks have been attached to the wall and put them back into the incubator.
5. Subsequently, the whole volume of fluid was changed every 3 days.

Results

After 3-5 days, cells could be seen crawling out around the tissue block, and 80-90% of the cells were fused after about 2 weeks.

Caveat

The final addition of 2 mL of culture solution in step 4 should flow slowly down the edge of the dish without the tissue mass, without blowing down the tissue mass.

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