PMID- 23845186
OWN - NLM
STAT- MEDLINE
DCOM- 20140501
LR  - 20181202
IS  - 1477-2566 (Electronic)
IS  - 1465-3249 (Linking)
VI  - 15
IP  - 9
DP  - 2013 Sep
TI  - Heparin concentration is critical for cell culture with human platelet lysate.
PG  - 1174-81
LID - S1465-3249(13)00516-1 [pii]
LID - 10.1016/j.jcyt.2013.05.006 [doi]
AB  - BACKGROUND AIMS: Culture media for mesenchymal stromal cells (MSCs) are generally 
      supplemented with fetal bovine serum. Human platelet lysate (hPL) has been proven 
      to be a very effective alternative without the risk of xenogeneic infections or 
      immune reactions. In contrast to fetal bovine serum, hPL comprises plasma, and 
      anticoagulants-usually unfractionated heparin (UFH)-need to be added to prevent 
      gel formation. METHODS: Cultures of MSCs in hPL media with various concentrations 
      of UFH and enoxaparin, a low-molecular-weight heparin (LMWH), were systematically 
      compared with regard to proliferation, fibroblastoid colony-forming unit 
      frequency, immunophenotype and in vitro differentiation. RESULTS: At least 0.61 
      IU/mL UFH or 0.024 mg/mL LMWH was necessary for reliable prevention of 
      coagulation of hPL pools used in this study. Higher concentrations impaired 
      cellular proliferation in a dose-dependent manner even without benzyl alcohol, 
      which is commonly added to heparins as a bacteriostatic agent. Colony-forming 
      unit frequency was also reduced at higher heparin concentrations, particularly 
      with LMWH, whereas no significant effect was observed on cellular morphology or 
      immunophenotype. High concentrations of heparins reduced the in vitro 
      differentiation toward adipogenic and osteogenic lineages. CONCLUSIONS: Heparin 
      concentration is critical for culture of MSCs in hPL media; this is of particular 
      relevance for cellular therapy where cell culture procedures need to be optimized 
      and standardized.
CI  - Copyright (c) 2013 International Society for Cellular Therapy. Published by 
      Elsevier Inc. All rights reserved.
FAU - Hemeda, Hatim
AU  - Hemeda H
AD  - Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical 
      School, Aachen, Germany.
FAU - Kalz, Jana
AU  - Kalz J
FAU - Walenda, Gudrun
AU  - Walenda G
FAU - Lohmann, Michael
AU  - Lohmann M
FAU - Wagner, Wolfgang
AU  - Wagner W
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130709
PL  - England
TA  - Cytotherapy
JT  - Cytotherapy
JID - 100895309
RN  - 0 (Cell Extracts)
RN  - 0 (Culture Media)
RN  - 9005-49-6 (Heparin)
SB  - IM
MH  - Adipogenesis/drug effects
MH  - Blood Platelets/*metabolism
MH  - Cell Differentiation/drug effects
MH  - Cell Extracts/*pharmacology
MH  - Cell Proliferation/drug effects
MH  - Cell Survival/drug effects
MH  - Cells, Cultured/*drug effects
MH  - Culture Media/*metabolism
MH  - Heparin/*pharmacology
MH  - Humans
MH  - Immunophenotyping/methods
MH  - Mesenchymal Stem Cells/drug effects
MH  - Osteogenesis/drug effects
OTO - NOTNLM
OT  - differentiation
OT  - fetal bovine serum
OT  - fibroblasts
OT  - human platelet lysate
OT  - mesenchymal stromal cells
OT  - platelet lysate gel
OT  - proliferation
OT  - serum
EDAT- 2013/07/13 06:00
MHDA- 2014/05/03 06:00
CRDT- 2013/07/13 06:00
PHST- 2012/11/15 00:00 [received]
PHST- 2013/04/25 00:00 [revised]
PHST- 2013/05/11 00:00 [accepted]
PHST- 2013/07/13 06:00 [entrez]
PHST- 2013/07/13 06:00 [pubmed]
PHST- 2014/05/03 06:00 [medline]
AID - S1465-3249(13)00516-1 [pii]
AID - 10.1016/j.jcyt.2013.05.006 [doi]
PST - ppublish
SO  - Cytotherapy. 2013 Sep;15(9):1174-81. doi: 10.1016/j.jcyt.2013.05.006. Epub 2013 
      Jul 9.