DOI: 10.5061/dryad.mk556
title: Data from: Influence of factors of cryopreservation and hypothermic storage on survival and functional parameters of multipotent stromal cells of placental origin

description: Human placenta is a highly perspective source of multipotent stromal cells
 (MSCs) both for the purposes of patient specific auto-banking and
 allogeneic application in regenerative medicine. Implementation of new GMP
 standards into clinical practice enforces the search for relevant methods
 of cryopreservation and short-term hypothermic storage of placental MSCs.
 In this paper we analyze the effect of different temperature regimes and
 individual components of cryoprotective media on viability, metabolic and
 culture properties of placental MSCs. We demonstrate (I) the possibility
 of short-term hypothermic storage of these cells; (II) determine DMSO and
 propanediol as the most appropriate cryoprotective agents; (III) show the
 possibility of application of volume expanders (plasma substituting
 solutions based on dextran or polyvinylpyrrolidone); (IV) reveal the
 priority of ionic composition over the serum content in cryopreservation
 media; (V) determine a cooling rate of 1°C/min down to -40°C followed by
 immersion into liquid nitrogen as the optimal cryopreservation regime for
 this type of cells. This study demonstrates perspectives for creation of
 new defined cryopreservation methods towards GMP standards.
description: Fig 1. Characterization
 of primary cell culture derived from placenta for MSC
 origin.(A) RT-PCR analysis for
 expression of typical MSC markers: cells provide a signal for CD90, CD73,
 and CD105 MSC while being negative for CD34 hematopoietic marker. (B)
 Minus RT control for RT-PCR. (C,D) Cells display strong signals for CD73
 and CD105 in flow cytometry analysis.Fig1.tiffFig 2. Differentiation potential of primary cell culture derived from placenta into lineages of MSC origin.MSCs were differentiated into (A) adipogenic (Oil Red O staining), (B) osteogenic (Von Kossa staining), and (C) chondrogenic (Alcian blue staining) lineages as a proof of principle for plasticity of MSCs. Cells expanded in regular MSC culture medium represent the negative control (bar = 50 µm).Fig2.tifFig 3. MSCs of placenta after storage at +20°C (A,B) and 4°C (C,D).Trypan blue staining, (A,C); phase contrast microscopy (B,D). Scale bar is 200 µm.Fig3.tifFig. 4. Parameters of placental MSCs after cryopreservation with different cooling rates.(A) - cell number in the sample; (B) - viable cells according to Trypan blue and 7 AAD staining; (C) - functional activity of cells in studied samples; (D) – adherence to plastic. *- significance to positive control p&lt;0.05.Fig4.tiffFig. 4. Parameters of placental MSCs after cryopreservation with different cooling rates:Data set for Fig.4PLOS ONE Pogozhykh (Fig_4).xlsTable 1. Characteristics of placental MSCs after short-term storage at different temperature conditionsData set for Table 1PLOS ONE Pogozhykh (Table_1).xlsTable 2. Cryopreservation of placental MSCs with various CPAs.Data set for Table 2PLOS ONE Pogozhykh (Table_2).xlsTable 3. Cryopreservation of placental MSCs with various cryoprotective media.Data set for Table 3PLOS ONE Pogozhykh (Table_3).xls