PMID- 31593692
OWN - NLM
STAT- MEDLINE
DCOM- 20200414
LR  - 20200414
IS  - 1090-2392 (Electronic)
IS  - 0011-2240 (Linking)
VI  - 91
DP  - 2019 Dec
TI  - Me(2)SO- and serum-free cryopreservation of human umbilical cord mesenchymal stem 
      cells using electroporation-assisted delivery of sugars.
PG  - 104-114
LID - S0011-2240(19)30059-8 [pii]
LID - 10.1016/j.cryobiol.2019.10.002 [doi]
AB  - Cryopreservation is the universal technology used to enable long-term storage and 
      continuous availability of cell stocks and tissues for regenerative medicine 
      demands. The main components of standard freezing media are dimethyl sulfoxide 
      (hereinafter Me(2)SO) and fetal bovine serum (FBS). However, for manufacturing of 
      cells and tissue-engineered products in accordance with the principles of Good 
      Manufacturing Practice (GMP), current considerations in regenerative medicine 
      suggest development of Me(2)SO- and serum-free biopreservation strategies due to 
      safety concerns over Me(2)SO-induced side effects and immunogenicity of animal 
      serum. In this work, the effect of electroporation-assisted pre-freeze delivery 
      of sucrose, trehalose and raffinose into human umbilical cord mesenchymal stem 
      cells (hUCMSCs) on their post-thaw survival was investigated. The optimal 
      strength of electric field at 8 pulses with 100 mus duration and 1 Hz pulse 
      repetition frequency was determined to be 1.5 kV/cm from permeabilization 
      (propidium iodide uptake) vs. cell recovery data (resazurin reduction assay). 
      Using sugars as sole cryoprotectants with electroporation, 
      concentration-dependent increase in cell survival was observed. Irrespective of 
      sugar type, the highest cell survival (up to 80%) was achieved at 400 mM 
      extracellular concentration and electroporation. Cell freezing without 
      electroporation yielded significantly lower survival rates. In the optimal 
      scenario, cells were able to attach 24 h after thawing demonstrating 
      characteristic shape and sugar-loaded vacuoles. Application of 10% Me(2)SO/90% 
      FBS as a positive control provided cell survival exceeding 90%. Next, high glass 
      transition temperatures determined for optimal concentrations of sugars by 
      differential scanning calorimetry (DSC) suggest the possibility to store samples 
      at -80  degrees C. In summary, using electroporation to incorporate cryoprotective sugars 
      into cells is an effective strategy towards Me(2)SO- and serum-free 
      cryopreservation and may pave the way for further progress in establishing 
      clinically safe biopreservation strategies for efficient long-term biobanking of 
      cells.
CI  - Copyright (c) 2019 Elsevier Inc. All rights reserved.
FAU - Mutsenko, Vitalii
AU  - Mutsenko V
AD  - Institute for Multiphase Processes, Leibniz University Hannover, Hannover, 
      Germany. Electronic address: mutsenko@imp.uni-hannover.de.
FAU - Barlic, Ariana
AU  - Barlic A
AD  - Educell Ltd, Trzin, Slovenia.
FAU - Pezic, Tamara
AU  - Pezic T
AD  - University of Ljubljana, Faculty of Electrical Engineering, Ljubljana, Slovenia.
FAU - Dermol-Cerne, Janja
AU  - Dermol-Cerne J
AD  - University of Ljubljana, Faculty of Electrical Engineering, Ljubljana, Slovenia.
FAU - Dovgan, Barbara
AU  - Dovgan B
AD  - Educell Ltd, Trzin, Slovenia; University of Ljubljana, Faculty of Electrical 
      Engineering, Ljubljana, Slovenia.
FAU - Sydykov, Bulat
AU  - Sydykov B
AD  - Institute for Multiphase Processes, Leibniz University Hannover, Hannover, 
      Germany.
FAU - Wolkers, Willem F
AU  - Wolkers WF
AD  - Institute for Multiphase Processes, Leibniz University Hannover, Hannover, 
      Germany.
FAU - Katkov, Igor I
AU  - Katkov II
AD  - Laboratory of the Amorphous State, Institute for Natural and Engineering 
      Sciences, Belgorod National Research University, Belgorod, Russia; MIP Vitronix, 
      ltd, Belgorod, Russia.
FAU - Glasmacher, Birgit
AU  - Glasmacher B
AD  - Institute for Multiphase Processes, Leibniz University Hannover, Hannover, 
      Germany.
FAU - Miklavcic, Damijan
AU  - Miklavcic D
AD  - University of Ljubljana, Faculty of Electrical Engineering, Ljubljana, Slovenia.
FAU - Gryshkov, Oleksandr
AU  - Gryshkov O
AD  - Institute for Multiphase Processes, Leibniz University Hannover, Hannover, 
      Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191005
PL  - Netherlands
TA  - Cryobiology
JT  - Cryobiology
JID - 0006252
RN  - 0 (Cryoprotective Agents)
RN  - 57-50-1 (Sucrose)
RN  - B8WCK70T7I (Trehalose)
RN  - N5O3QU595M (Raffinose)
RN  - YOW8V9698H (Dimethyl Sulfoxide)
SB  - IM
MH  - Animals
MH  - Biological Specimen Banks
MH  - Cell Survival/drug effects
MH  - Cryopreservation/*methods
MH  - Cryoprotective Agents/*metabolism/*pharmacology
MH  - Dimethyl Sulfoxide/*pharmacology
MH  - Electroporation/*methods
MH  - Freezing
MH  - Humans
MH  - Mesenchymal Stem Cells/*cytology
MH  - Raffinose/metabolism/pharmacology
MH  - Sucrose/metabolism/pharmacology
MH  - Tissue Engineering
MH  - Trehalose/metabolism/pharmacology
MH  - Umbilical Cord/cytology
OTO - NOTNLM
OT  - Electroporation
OT  - Human umbilical cord mesenchymal stem cells
OT  - Me(2)SO- and serum-free cryopreservation
OT  - Permeabilization
OT  - Sugars
EDAT- 2019/10/09 06:00
MHDA- 2020/04/15 06:00
CRDT- 2019/10/09 06:00
PHST- 2019/05/02 00:00 [received]
PHST- 2019/08/23 00:00 [revised]
PHST- 2019/10/03 00:00 [accepted]
PHST- 2019/10/09 06:00 [pubmed]
PHST- 2020/04/15 06:00 [medline]
PHST- 2019/10/09 06:00 [entrez]
AID - S0011-2240(19)30059-8 [pii]
AID - 10.1016/j.cryobiol.2019.10.002 [doi]
PST - ppublish
SO  - Cryobiology. 2019 Dec;91:104-114. doi: 10.1016/j.cryobiol.2019.10.002. Epub 2019 
      Oct 5.