23 Jul METHOD FOR PREPARING TISSUE-SPECIFIC MATRIX FOR TISSUE ENGINEERING OF PARENCHYMAL ORGAN
Abstract
FIELD: medicine.SUBSTANCE: tissue-specific matrix is obtained by performing a perfusion washing and a decellularisation of a parenchymal organ. Herewith 60-120 minutes before the perfusion washing of the donor organ, measures to prevent intravascular blood cell aggregation and microcirculation disturbances are taken and involve the intramuscular or intravenous administration of a disaggregant or disaggregants (heparin and trental) into a donor. That is followed by an organ perfusion by administering into its vascular bed phosphate-buffered normal saline of the following composition: 138 mM NaCl, 2.67 mM KCl, 1.47 mM KHPO, 8.1 mM NaHPO, distilled water up to 1l and containing 1% serum albumin and 10-15% glycerol or 10-15% dimethyl sulphoxide with pH 7.4 in an amount equal to a double amount of the vascular bed of the organ at a perfusion pressure of 100-120 mm Hg in its arterial system. Thereafter, the decellularised organ is ground to a particle size of 0.5mm to 4mm; the ground fragments are portioned in an amount of 5-10g, and each portion is frozen by immersing into liquid nitrogen gas for 5-10 minutes. The frozen portions are re-ground to a particle size of no more than 600 mcm; then each portion is de-frozen by re-suspending in phosphate-buffered normal saline 30-70ml of the following composition containing 138 mM NaCl, 2.67 mM KCl, 1.47 mM KHPO, 8.1 mM NaHPO, distilled water up to 1l , with pH 7.4, at a room temperature. The prepared suspension is cleaned from the particles of less than 200mcm; the prepared fraction is lyophilised and sterilised to prepare the target matrix samples.EFFECT: using the invention enables providing the more complete decellularisation procedure ensured by preventing the disturbed microcirculation in the donor organ, providing higher density and uniformity of the re-cellularisation of the entire matrix and easier control thereof by increasing the matrix adhesive surface areas for the contact to inoculating cells as prepared in the form of powder.4 cl, 5 ex, 9 dwg
Year: 2015
Country: RU
Doc No: 2539918