Tissue engineering is an approach to repair damaged or defected tissue with stem cells and scaffolds. Stem cells used in tissue engineering can be easily expanded in vitro, and can be differentiate to other cell types. To secure the various sources of stem/progenitor cells, stromal cells from various tissues were isolated and characterized. Bone marrow stormal cells (BMSCs), also known as mesenchymal stem cells, can differentiate into other cell types and they are best in their utilization potential among many skeletal stem cells. Various musculoskeletal tissues such as adipose, xiphisternum cartilage, tendon, and ligament contain stromal cells, which can be isolated using collagenase type I. Isolated cells were named as Adipose derived stem cells (ADSCs), xiphisternum derived stem cells (XDSCs), tendon derived stem cells (TDSCs), and ligament derived stem cells. Although it is probable that every isolated stem cells displayed different characteristics, there has been no approach to compare those cells in exactly same conditions. In this study, various tissue derived stem cells were comparatively characterized for the cell morphology, colony morphology, CFU-F, and surface marker expressions. Stem cells derived from various tissues displayed morphology of fibroblast-like shape, although there were subtle differences between the skeletal stem cells according to their origin. Based on CFU-F assay, all the cells showed density-independent growth when they were seeded in clonogenic density. All the stem cells tested showed positive expression of CD90 and α-SMA and negative to CD45 displaying similar characteristics to BMSCs.