Abstract
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.
Key Words
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Article info
Publication history
Published online: July 11, 2013
Accepted:
May 11,
2013
Received:
November 15,
2012
Identification
Copyright
© 2013 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
