Comparison of cultured mesenchymal stem cells derived from bone marrow or peripheral blood of rats
Author: Achmad F. Kamal, Diah Iskandriati, Ismail H. Dilogo, Nurjati C. Siregar, Errol U. Hutagalung, Achmad A. Yusuf, Silmi Mariya, Kurniadi HusodoAbstract
Objectives: Mesenchymal stem cells (MSC) are primarily isolated from bone marrow. Peripheral blood is also reported as an alternative source of MSC. This study compared MSC which were isolated and cultured from bone marrow and those from peripheral blood of rats.
Methods: MSC from bone marrow and peripheral blood were harvested from 5 male Sprague Dawley rats. After isolation, the cells were grown on tissue culture plates with concentration of 107 cells per well. Observations were conducted to evaluate the attachment of nucleated cells with fibroblast-like morphology. Characterization of MSC was done using reverse transcriptasepolymerase chain reaction (RT-PCR) and immunocytochemistry assay.
Results:The mean number of nucleated cells isolated from the bone marrow on day 0 was higher Than those isolated from the peripheral blood. Bone marrow MSC with typical fibroblast-like morphology proliferated rapidly and reached 80% confluency on day 14. Subcultures were able to be conducted on day 15 (first passage) and day 28 (second passage). On days 15 and 28, no nucleated cells remained in peripheral blood cultures.
Conclusions: Bone marrow derived MSC are more enriched and grow more efficiently than those derived from peripheral blood.
Key Words: Bone marrow; Culture; Mesenchymal stem cells; Peripheral blood
