Elaborating of a cryoconservation protocol of blood vessels for the tissue-specific matrices
Heading: radiation medicine Article type: Original article
Authors: Lauk-Dubitsky S.E., Astrelina Т.А., Brumberg V.A., Nikitina V.A., Suchkova Yu.B., Usupzhanova D.Yu., Brunchukov V.A., Rastorgueva A.A., Lomonosova Е.Е., Kobzeva I.V., Makhova A.E., Karaseva T.V., Bushmanov A.Yu., Samoilov A.S.
Organization: State Scientific Research Center n.a. A.I. Burnasyan — Federal Medical Biophysical Center of Federal Medical Biological Agency
Purpose: to create a protocol for complex cryoconservation of blood vessels in polydimethylsiloxane to create tissue-specific matrices. Material and Methods. The material ofthe study was the iliac arteries from postmortem donors, seized in the multi-organ donation, rejected for unclaimed and disposed of later. Polydimethylsiloxane was used as a coolant (PDMS) with a viscosity of 5-25 centistokes (Spectroplast, Russia). Bench testing of cooling and heating ofthe blood vessel in PDMS, testing of heating and cooling of PDMS up to operating temperatures (-80°C) in different ways and variants of its thermal insulation, assessment of different cooling rates ofthe vessel during cryoconservation, modeling of 3 types of working chambers for cryoconservation, sterilization and defrosting of up to 10 vessels, modeling of cryoconservation and thawing of vessels in PDMS under different conditions, selection of substrates from silicone and plastic for uniform fixation of vessels which are of size or size in the working chambers or without them. Results. The developed protocol allows cryopreserved from 1 to 10 or more vessels of all basic sizes both immediately after removal and after transportation to the laboratory, to vary the cooling rate from 1.5 to 300°C/min and above, to be used with different levels of laboratory equipment, as with liquid nitrogen, dry ice, and only with a medical freezer. Conclusion. The developed protocol allows to apply it in cases of complex cryoconservation of blood vessels to create tissue-specific matrices and can be recommended for preclinical testing.
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