Abstract
BackgroundIn vivo tissue regeneration depends on migration of stem cells into injured areas, their differentiation into specific cell types, and their interaction with other cells that are necessary to generate new tissue. Human mesenchymal stem cells, a subset of bone marrow stromal cells (BMSCs), can migrate and differentiate into osteoblasts in bone tissue. This can be facilitated by recombinant growth factors and cytokines. In many animal species, the availability of genomic sequences, recombinant proteins, and/or antibodies is limited so that new approaches are needed to generate resources that facilitate migration of stem cells into tissue defect areas. Here we used bone marrow stromal cells of human, ovine, equine, and canine origin to generate hypoxia-conditioned media (HCM) in order to attract BMSCs of the respective species in migration assays.ResultsWe show that HCM contain attractors even more potent than vascular endothelial growth factor and can therefore be used in many animal species without the need for purified proteins.ConclusionGeneration of HCM is easy and cheap compared to preparation and purification of protein fractions and/or recombinant proteins. Hence, HCM could be applied in large animals (e.g. sheep, horse, dogs) for attraction of BMSCs into tissue defects caused by tumor resection or trauma.
Highlights
In vivo tissue regeneration depends on migration of stem cells into injured areas, their differentiation into specific cell types, and their interaction with other cells that are necessary to generate new tissue
vascular endothelial growth factor (VEGF) and HMGB1 enzyme-linked immunosorbent assay (ELISA) of human, ovine, equine, and canine hypoxia-conditioned media (HCM) We performed ELISAs in order to determine the amount of VEGF and HMGB1 released from bone marrow stromal cells (BMSCs) under hypoxic conditions
When we started our experiments, only human VEGF kit was available; we tested if ovine, equine, or canine VEGF could be detected by antibodies directed against human VEGF by using human, ovine, equine, and canine HCM
Summary
In vivo tissue regeneration depends on migration of stem cells into injured areas, their differentiation into specific cell types, and their interaction with other cells that are necessary to generate new tissue. A subset of bone marrow stromal cells (BMSCs), can migrate and differentiate into osteoblasts in bone tissue. This can be facilitated by recombinant growth factors and cytokines. The availability of genomic sequences, recombinant proteins, and/or antibodies is limited so that new approaches are needed to generate resources that facilitate migration of stem cells into tissue defect areas. We used bone marrow stromal cells of human, ovine, equine, and canine origin to generate hypoxia-conditioned media (HCM) in order to attract BMSCs of the respective species in migration assays. We used BMSCs of human, ovine, equine, and canine origin to generate hypoxia-conditioned media (HCM) in order to attract BMSCs of the respective species in migration assays. We were able to show the presence of VEGF and high-mobility group protein
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