Multi-potent differentiation of human purified muscle-derived cells: potential for tissue regeneration.

Abstract

To investigate whether CD34+ purified human muscle-derived cells (hMDCs) are capable of multiple lineage differentiation. The hMDCs were isolated from human skeletal muscle and purified using a CD34+ cell selection system (Dynal Biotech, Oslo, Norway). Adherent populations of cells were expanded in culture and cell differentiation was induced using different kinds of growth factors and different differentiation-conditional media. The immunohistochemical properties of CD34+ hMDCs were examined after varying periods in culture. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to investigate the gene expression of the undifferentiated and differentiated hMDCs. Using special differentiation conditions the CD34+ hMDCs could be differentiated into myogenic cells, adipocytes, osteocytes and chondrocytes. The differentiation was confirmed by immunohistochemistry. RT-PCR and Western blotting showed multiple-lineage gene-level expression in the different cultivation periods of the differentiated cells. We confirmed the multi-lineage capacity of a population of stem cells, termed CD34+ hMDCs. Our findings showed that CD34+ hMDCs are capable of multiple mesodermal-lineage differentiation, as shown by the expression of several lineage-specific genes. They can be differentiated toward the myogenic, osteogenic, adipogenic and chondrogenic lineages. These cells might have potential for use in tissue regeneration.