ABSTRACT
Background: Although bone marrow serves as the ‘gold standard’ MSC source, adipose tissue has become a promising alternative source. Passage and cryopreservation are two effective methods to multiply, pool, and store MSC without altering its function Aims: To investigate the passage effects on the senescence profile of cryopreserved bone marrow and adipose-derived mesenchymal stem cells (MSCs). Study Design: Analytical observational study. Place and Duration of Study: Stem Cell Medical Technology Integrated Service Unit, Faculty of Medicine, Universitas Indonesia—Cipto Mangunkusumo Hospital, Jakarta, Indonesia, during the period of April to September 2016. Methodology: We analyzed the viability, cell size, population doubling time (PDT), percentage of senescent cells, and colony forming unit. Samples were bone marrow and adipose MSCs at passage one, which was cryopreserved for the first and second time. Numerical data were analyzed using the Student’s T test and analysis of variance (ANOVA) test. Results: Both in once and twice cryopreservation group, PDT and senescent cell percentage of bone marrow and adipose tissue MSCs differed significantly, where the PDT senescent cell percentage values of bone marrow MSCs were higher in all passages compared to adipose tissue. Regarding 30% confluence cell size and viability, significant differences between once and twice cryopreservation group varied and did not show any trend. The cell size and viability were less 2500 µm2, and more than 85%, respectively. Therefore, the difference in cell size at 30% confluence and viability might be regarded as normal variations. Conclusion: Cryopreserved adipose-derived MSCs showed better results compared to cryopreserved bone marrow-derived MSCs in terms of population doubling time (PDT) and senescence.
ABSTRACT
Background: Red marrow has been described as the main source of mesenchymal stem cells although its aspiration and isolation from bone marrow was reported to have significant donor site morbidity. Since secondary bone healing occurs through formation of callus as the result of proliferation and differentiation of mesenchymal stem cells, callus may become alternative source for mesenchymal stem cells. In this study, we compared the number of plastic-adherent cells from fracture site callus and bone marrow of iliac crest after two and four weeks of culture. Methods: Sixteen New Zealand rabbits were fracturized at the femoral shaft. Then, these rabbits were taken care. After two weeks of fracturization, 3 mL iliac crest bone marrow aspiration and callus extraction of eight rabbits were cultured (group I). The other eight rabbits were treated equally after four weeks of fracturization (group II). Simultaneously, the cultures were observed after one and two weeks. Four weeks later, they were harvested. Cells were counted using Neubauer hemocytometer. The average number of cells between the sources and groups were statistically analyzed using the unpaired t-test. Results: In group I, there were 2.6 ± 0.1 x 104 cells in the culture of iliac crest bone marrow aspirate and 2.5 ± 0.1 x 104 cells in culture of callus extract from fracture site (p = 0.34). In group II, there were 2.7 ± 0.1 x 104 cells and 2.1 ± 0.1 x 104 cells, respectively (p < 0.001). Conclusion: Fracture site callus at the second week post-fracturization may be potential as source of plastic-adherent cells compared with iliac crest bone marrow.