PMID- 18765238
OWN - NLM
STAT- MEDLINE
DCOM- 20090122
LR  - 20151119
IS  - 1090-2392 (Electronic)
IS  - 0011-2240 (Linking)
VI  - 57
IP  - 3
DP  - 2008 Dec
TI  - Cryopreservation of human fetal liver hematopoietic stem/progenitor cells using 
      sucrose as an additive to the cryoprotective medium.
PG  - 195-200
LID - 10.1016/j.cryobiol.2008.08.003 [doi]
AB  - INTRODUCTION: Human fetal liver (HFL) is a valuable source of hematopoietic 
      stem/progenitor cells (HSCs) for the treatment of various hematological 
      disorders. This study describes the effect of sucrose addition to a 
      cryoprotective medium in order to reduce the Me(2)SO concentration during 
      cryopreservation of HFL hematopoietic cell preparations. METHODS: Human fetal 
      liver (HFL) cells of 8-12 weeks of gestation were cryopreserved with a cooling 
      rate of 1 degrees C/min down to -80 degrees C and stored in liquid nitrogen. The 
      cryoprotectant solutions contained 2% or 5% Me(2)SO (v/v) with or without sucrose 
      at a final concentration of 0.05, 0.1, 0.2 or 0.3M. The metabolic activity of HFL 
      cells was determined using the alamar blue assay. For the determination of the 
      number and survival of hematopoietic progenitors present, cells were stained with 
      CD34 (FITC) and 7-AAD, and analyzed by flow cytometry. The colony-forming 
      activity of HFL hematopoietic stem/progenitor cells after cryopreservation was 
      assessed in semisolid methylcellulose. RESULTS: The addition of sucrose to the 
      cryoprotective medium produced a significant reduction in HFL cell loss during 
      cryopreservation. The metabolic activity of HFL cells, cryopreserved with 5% 
      Me(2)SO/0.3M sucrose mixture was comparable to cryopreservation in 5% Me(2)SO/10% 
      FCS. Although the inclusion of sucrose did not affect the survival of CD34(+) 
      cells in HFL after cryopreservation it did improve the functional capacity of 
      hematopoietic stem/progenitor cells. CONCLUSION: The inclusion of sucrose as an 
      additive to cryoprotective media for HFL cells enables a reduction in the 
      concentration of Me(2)SO, replacing serum and increasing the efficiency of 
      cryopreservation.
FAU - Petrenko, Yu A
AU  - Petrenko YA
AD  - Institute for Problems of Cryobiology and Cryomedicine of NASU, Kharkov 61015, 
      Ukraine.
FAU - Jones, D R E
AU  - Jones DR
FAU - Petrenko, A Yu
AU  - Petrenko AY
LA  - eng
GR  - 069472/Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080814
PL  - Netherlands
TA  - Cryobiology
JT  - Cryobiology
JID - 0006252
RN  - 0 (Oxazines)
RN  - 0 (Xanthenes)
RN  - 1FN9YD6968 (resazurin)
RN  - 57-50-1 (Sucrose)
RN  - YOW8V9698H (Dimethyl Sulfoxide)
SB  - IM
MH  - Cell Survival
MH  - Cells, Cultured
MH  - Colony-Forming Units Assay
MH  - Cryopreservation/*methods
MH  - Dimethyl Sulfoxide/pharmacology
MH  - Hematopoietic Stem Cells/cytology/*drug effects/metabolism
MH  - Humans
MH  - Liver/cytology/embryology
MH  - Oxazines/metabolism
MH  - Sucrose/*pharmacology
MH  - Xanthenes/metabolism
EDAT- 2008/09/04 09:00
MHDA- 2009/01/23 09:00
CRDT- 2008/09/04 09:00
PHST- 2007/08/22 00:00 [received]
PHST- 2008/04/25 00:00 [revised]
PHST- 2008/08/05 00:00 [accepted]
PHST- 2008/09/04 09:00 [pubmed]
PHST- 2009/01/23 09:00 [medline]
PHST- 2008/09/04 09:00 [entrez]
AID - S0011-2240(08)00110-7 [pii]
AID - 10.1016/j.cryobiol.2008.08.003 [doi]
PST - ppublish
SO  - Cryobiology. 2008 Dec;57(3):195-200. doi: 10.1016/j.cryobiol.2008.08.003. Epub 
      2008 Aug 14.