Phenotypical and functional characteristics of mesenchymal stem cells from bone marrow: comparison of culture using different media supplemented with human platelet lysate or fetal bovine serum

Abstract

IntroductionMesenchymal stem cells (MSCs) are multipotent cells able to differentiate into several mesenchymal lineages, classically derived from bone marrow (BM) but potentially from umbilical cord blood (UCB). Although they are becoming a good tool for regenerative medicine, they usually need to be expanded in fetal bovine serum (FBS)-supplemented media. Human platelet lysate (HPL) has recently been proposed as substitute for safety reasons, but it is not yet clear how this supplement influences the properties of expanded MSCs.MethodsIn the present study, we compared the effect of various media combining autologous HPL with or without FBS on phenotypic, proliferative and functional (differentiation, cytokine secretion profile) characteristics of human BM-derived MSCs.ResultsDespite less expression of adipogenic and osteogenic markers, MSCs cultured in HPL-supplemented media fully differentiated along osteoblastic, adipogenic, chondrogenic and vascular smooth muscle lineages. The analyses of particular specific proteins expressed during osteogenic differentiation (calcium-sensing receptor (CaSR) and parathormone receptor (PTHR)) showed their decrease at D0 before any induction for MSC cultured with HPL mostly at high percentage (10%HPL). The cytokine dosage showed a clear increase of proliferation capacity and interleukin (IL)-6 and IL-8 secretion.ConclusionsThis study shows that MSCs can be expanded in media supplemented with HPL that can totally replace FBS. HPL-supplemented media not only preserves their phenotype as well as their differentiation capacity, but also shortens culture time by increasing their growth rate.

Placental Mesenchymal Stromal Cells Derived from Blood Vessels or Avascular Tissues: What Is the Better Choice to Support Endothelial Cell Function?

“…A reason for this could be that bv-MSCs, in contrast to av-MSCs, secreted noticeable amounts of VEGF, an important inducer of angiogenesis, as shown by quantitative ELISA analysis. While MSCs from bone marrow are known to secrete VEGF [45][46][47][48], its secretion by av-MSCs has not been reported before, which is in accordance with our results. As less VEGF was found in the Cdm of av-MSCs compared with the control medium, we speculate that these cells take up VEGF from the medium and metabolize it in a similar extent as ECs.…”

Tumour-associated fibroblasts contribute to neoangiogenesis in human parathyroid neoplasia

“…We investigated the effect of the potent CASR agonist R568 on VEGFA mRNA levels in hBM-MSCs, as these cells have been demonstrated to express functional CASR, possibly involved in osteoblast differentiation (Ben Azouna et al 2012). Treatment of hBM-MSCs with R568 (50 mM) for 5 days in presence of 1.5 mM extracellular calcium, a condition previously shown to activate CASR in cultured human parathyroid cells (Terranegra et al 2010), induced a significant reduction in PTH mRNA levels in PAd-derived cells, as expected, and of VEGFA mRNA levels in hBM-MSCs.…”

Standardization and Safety of Alveolar Bone–derived Stem Cell Isolation

“…The average population-doubling time (PDT) was calculated between passage 1 (P1) and passage 3 (P3) as t/n, where t is the duration of culture in days and n is the number of population doublings (PD), calculated according to the formula n = (log N h -log N i )/log2 (Ben Azouna et al, 2012). N h denotes the number of cells harvested at the time of counting at the end of a passage prior to subculture, and N i refers to the number of cells initially plated at the beginning of the passage.…”