Abstract
Adipose tissue is a promising source of mesenchymal stem cells. Their potential to differentiate and regenerate other types of tissues may be affected by several factors. This may be due to in vitro cell-culture conditions, especially the supplementation with antibiotics. The aim of our study was to evaluate the effects of a penicillin-streptomycin mixture (PS), amphotericin B (AmB), a complex of AmB with copper (II) ions (AmB-Cu2+) and various combinations of these antibiotics on the proliferation and differentiation of adipose-derived stem cells in vitro. Normal human adipose-derived stem cells (ADSC, Lonza) were routinely maintained in a Dulbecco’s Modified Eagle Medium (DMEM) that was either supplemented with selected antibiotics or without antibiotics. The ADSC that were used for the experiment were at the second passage. The effect of antibiotics on proliferation was analyzed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and sulforhodamine-B (SRB) tests. Differentiation was evaluated based on Alizarin Red staining, Oil Red O staining and determination of the expression of ADSC, osteoblast and adipocyte markers by real-time RT-qPCR. The obtained results indicate that the influence of antibiotics on adipose-derived stem cells depends on the duration of exposure and on the combination of applied compounds. We show that antibiotics alter the proliferation of cells and also promote natural osteogenesis, and adipogenesis, and that this effect is also noticeable in stimulated osteogenesis.
Highlights
Adipose tissue is an excellent source of adult stem cells, which are called adipose-derived stem cells (ADSC) [1]
To gain better understanding of the used experimental systems in regenerative medicine and tissue engineering, we have evaluated the effect of antibiotics on the growth and differentiation of the MSC derived from the adipose tissue commonly used in cell culture settings and in the clinic
This study showed that amphotericin B and its new form with copper ions did produce significant effects of the osteogenesis, especially the lower expression of RUNX2, which is an early marker of osteogenesis and the osteopontin (SPP1) gene, which is a late marker of osteogenesis
Summary
Adipose tissue is an excellent source of adult stem cells, which are called adipose-derived stem cells (ADSC) [1] They hold great promise in cell replacement therapies, and studies demonstrated their properties in the regeneration of bones, joints or cardiomyocytes in vivo and in vitro [2,3,4]. Stem cells must be isolated and cultured under the optimal conditions for a particular cell type, in order to use them in transplantology. They have to be maintained in an undifferentiated state to preserve their self-renewal potential. It has been suggested that the antibiotics in a cell culture may change the regenerative potential and other biologic properties in many types of cells [13]
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