Obtainment of mesenchymal stem cells by surgeons

Summary Biological and clinical interest in mesenchymal stem cells (MSCs) has risen dramatically over the past two decades. This paper introduces the methods we used to obtain bone marrow MSCs by means of locally available materials. Rabbit bone marrow contents were harvested from rabbit femurs. After lysing red blood cells, the fibroblast-like cells adhering to the flask were collected. The obtained cells from Passage 3 were tested for the potential to differentiate into mesenchymal tissues (osteocytes, chondrocytes, and adipocytes). The cells were also tested for the presence of marker proteins of mesenchymal lineages and the absence of marker proteins of hematopoietic lineages. All of the materials we used to induce differentiation and characterization of cells was obtained from local biotechnical companies. The cells obtained showed evidence of proliferation. After in vitro correlative induction, the cells showed the presence of calcium deposition, type II collagen formation, and intracellular accumulation of lipid vacuoles. The cells were also proven to have the presence of a marker protein of mesenchymal lineages (CD44) and the absence of marker proteins of hematopoietic lineages (CD14 and CD45). It is concluded that surgeons can easily obtain MSCs using locally available materials. We would like to share our experience with those who are interested in the research of MSCs.