Abstract
Authors studied potential side effects of fetal calf serum (FCS) in cultivation media on human dental pulp stem cells (DPSC) during long-term cultivation. Two lines of DPSC obtained healthy donors (male 22 years, female 23 years) were used. Both lines were cultivated under standard cultivation conditions in four different media containing 10% or 2% FCS and substituted with growth factors. During long-term cultivation proliferation ability, karyotype and phenotype of DPSC were measured. Both lines of DPSC cultivated in a media containing 2% FCS and ITS supplement showed the highest number of population doublings. On the other hand the proliferation rate of DPSC cultivated in a media with 2% FCS without ITS supplement was slowest. Proliferation rate of DPSC cultivated in 10% FCS media with or without FGF-2 was comparable. DPSC cultivated in a media with 10% FCS showed a significantly higher amount of chromosomal aberrations. These chromosomal aberrations do not seem to be clonal but surprisingly we found large amounts of tetraploid cells in the 9th passage in both media containing 10% FCS. Our study proved that cultivation of DPSC in media containing higher concentration of FCS has critical side effects on cell chromosomal stability.
Highlights
Multipotent mesenchymal stromal stem cells (MSC) are defined as cells which are able to selfrenew or differentiate into other mature cell types
In this study we have focused on finding how the fetal calf serum (FCS) can affect dental pulp stem cells (DPSC) and which supplements could be used in the case of decreasing amount of FCS
Media A composed of alfa-MEM, 2% FCS, 10 ng/ml EGF, 10 ng/ml PDGF, 0.2 mM L-ascorbic acid, 2% glutamine, 100 U/ml penicilin/ 100 μg/ml streptomycin (Gibco, UK), 20 μg/ml gentamycin, 50 nM dexamethasone (Sigma, USA) and 10 μl/ml ITS is our standard cultivation media and DPSC cultivated in this media were used as control group
Summary
Multipotent mesenchymal stromal stem cells (MSC) are defined as cells which are able to selfrenew or differentiate into other mature cell types (at last into adipocytes, chondrocytes and ostoblasts). These cells are able to adhere to plastic surface of the cultivation flasks, are supposed to be positive at least for CD105, CD73, CD90 and have imunosupressive abilities and can undergo gene modification. In contrast with embryonic stem cells, isolation of MSC does not provoke any ethical issues For these reasons, these cells promised to be a good source for tissue engineering and gene therapy
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