Author(s)

Zhang Yu ¹, Wang Jing ¹

Affiliation(s)

¹ Hebei North University, 075000, Zhangjiakou, China

Corresponding Author

Zhang Yu

ABSTRACT

Lymphatic smooth muscle cells are the basis of the study of lymphatic microcirculation. Traditional lymphatic culture often uses patching and enzymatic hydrolysis. However, because the traditional enzymatic hydrolysis method has a low success rate in culturing cells, this article introduces the use of trypsin digestion to treat rat thoracic ducts. Culture the smooth muscle cells of the lymphatic vessels, and compare the advantages and disadvantages of trypsin digestion and other smooth muscle cell culture methods. Experimental operation: select 4 fasting male wistar rats, anesthetize the rats by intraperitoneal injection of 1% sodium pentobarbital, and soak them in 75% alcohol for 5 minutes for disinfection. Place the rat on a sterile table for thoracotomy, take out the thoracic duct and surrounding tissues, quickly put in PSS cold solution for later use, separate the rat's thoracic lymphatic vessels under a microscope, rinse with PSS 3 times in the cold night to remove the floating liquid . Add warm DMEM to 37°C water bath and incubate for 30min, discard DMEM, add 2ml Trypsin and put it in 37°C water bath for digestion for 10min (vibrate once every 2min). Aspirate the digestion solution and wash it with DPBS for 3 times. Take out the thoracic duct and cut it into pieces. Add 4ml collagenase and put it in a 37°C water bath for 30min (vibrate once every 2min). After digestion, add 25%D/F2 4ml Terminate the digestion, and then perform centrifugation (rotating speed 1100r/min centrifugation for 3 minutes). Finally, the cells were resuspended in 2ml of 25% special culture medium, and 5% CO was injected into the cells. They were cultured in a 37°C incubator and regularly observed under the microscope.

KEYWORDS

Cell biology, Trypsin digestion method to culture smooth muscle cells, Cell culture, Thoracic aortic lymphatic vessels, Lymphatic smooth muscle cells, Rats

CITE THIS PAPER

Zhang Yu, Wang Jing. Culture of Rat Lymphatic Smooth Muscle Cells. MEDS Clinical Medicine (2021) 2: 58-60. DOI: http://dx.doi.org/10.23977/medsc.2021.020111.

REFERENCES

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