23 Jul METHOD OF DECELLULARISATION OF SMALL-CALIBRE BLOOD VESSELS
Abstract
FIELD: medicine.SUBSTANCE: invention relates to medicine, namely to regenerative medicine and tissue engineering, and can be applied for obtaining extracellular matrixes of small-caliber blood vessels. For this purpose at the first stage of tissues fragment of blood vessel is washed with distilled water for 1 hour at temperature +4°C. After that, fragment is placed into 0.05% solution of tripsin and 0.02% EDTA for 1 hour at temperature +37°C. At the third stage processing in 0.075% solution of sodium dodecylsulfate is performed for 24 hours at temperature 26°C. After that, fragment is placed into 0.25% solution of Triton X-100 for 24 hours at temperature 26°C. At the fourth stage said fragment is processed with solution which contains 20 mcg/ml of RNA-ase and 200 mcg/ml of DNA-ase I, for 6 hours at temperature +37°C. After each stage of processing, fragment of blood vessel is three times washed in phosphate-salt buffer solution, for 10 minutes each time. The entire process of processing is carried out with constant mixing of solutions and simultaneous vibration, created by vibromotor, located on the external wall of the reservoir.EFFECT: method makes it possible to increase quality of decellularisation of small-calibre blood vessels, preserve their integrity and ultrastructure for further immobilisation of recipient’s cells.2 ex, 6 dwg
Year: 2014
Country: RU
Doc No: 2504334