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Theory-based cryopreservation mode of mesenchymal stromal cell spheroids.
Gordiyenko, O I; Kovalenko, I F; Rogulska, O Y; Trufanova, N A; Gurina, T M; Trufanov, O V; Petrenko, O Y.
Afiliación
  • Gordiyenko OI; Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine. Electronic address: olga.gordiyenko.ipcic@gmail.com.
  • Kovalenko IF; Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine. Electronic address: i.kovalenko.ipcic@gmail.com.
  • Rogulska OY; Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine; Institute of Physiology, Czech Academy of Science, Prague, Czech Republic; Institute of Experimental Medicine, Czech Academy of Science, Prague, Czech Republic. Electronic address: rogu
  • Trufanova NA; Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine. Electronic address: n.a.trufan@gmail.com.
  • Gurina TM; Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine. Electronic address: gurtata@ukr.net.
  • Trufanov OV; Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine. Electronic address: olegtrufanov2015@gmail.com.
  • Petrenko OY; Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine. Electronic address: alexander_petrenko@cryo.org.ua.
Cryobiology ; 115: 104906, 2024 Jun.
Article en En | MEDLINE | ID: mdl-38762155
ABSTRACT
Cryopreservation of spheroids requires development of new improved methods. The plasma membranes permeability coefficients for water and cryoprotectants determine time characteristics of mass transfer through the cell membranes, and therefore the optimal modes of cells cryopreservation. Here we proposed an approach to cryopreservation of multicellular spheroids which considers their generalized characteristics as analogues of the membranes' permeability coefficients of the individual cells. We have determined such integral characteristics of spheroids from mesenchymal stromal cells (MSCs) as osmotically inactive volume; permeability coefficients for water and Me2SO molecules and the activation energy of their penetration. Based on these characteristics, we calculated the osmotic behavior of multicellular spheroids under cooling conditions to select the optimal cooling rate. We also determined the optimal cooling rate of spheroids using the probabilistic model developed based on the two-factor theory of cryodamage. From the calculation it follows that the optimal cooling rate of the MSC-based spheroids is 0.75°Ð¡/min. To verify the obtained theoretical estimates, we conducted experiments on freezing MSC-based spheroids under different modes. The obtained results of primary viability screening indicate that freezing at a constant linear cooling rate of 0.75-1.0°Ð¡/min gives a good result. Theoretical prediction of the spheroid osmotic behavior during cooling provided the basis for experimental verification of varying the temperature to which slow cooling should be carried out before immersion in liquid nitrogen. Slow freezing of spheroids to -40 °C followed by immersion in liquid nitrogen was shown to preserve cells better than slow freezing to -80 °C. Obtained data allow more effective use of MSC-based spheroids in drug screening and regenerative medicine.
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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Criopreservación / Supervivencia Celular / Esferoides Celulares / Crioprotectores / Células Madre Mesenquimatosas Límite: Humans Idioma: En Revista: Cryobiology Año: 2024 Tipo del documento: Article

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Criopreservación / Supervivencia Celular / Esferoides Celulares / Crioprotectores / Células Madre Mesenquimatosas Límite: Humans Idioma: En Revista: Cryobiology Año: 2024 Tipo del documento: Article