Cellularization of small sized grafts from biological material using the gravitational modality principle

Abstract

The research aims to present an improved cellularization process of small sized grafts obtained from biological materials through utilisation of gravitational modality principle. The obvious problem of the biological materials cellularization process, is failure in instant absorption of poured cellular suspension, as a result it leads to cells loss because of suspension spreading through the used dishes, which is especially characteristic for small sized materials. For transplantation were cellularized grafts obtained from biological materials of 44,31mm 3±2,72mm 3, with chondrocytes (n=18) and mesenchymal stem cells (MSC) (n=20). With chondrocytes in a well of 96-well microplates (Mp96) were cellularized 7 grafts with 2.8x106±2.38x105 cells/graft and in DFCSSG –11 grafts with 2.87x106±3x105 cells/graft (p˃0.5). With MSC in Mp96 were cellularized 8 grafts with 1.29x106±2x105 cells/graft and in DFCSSG –12 grafts with 1.41x106±3.29x105 cells/graft (p˃0.1). All cellularized grafts were held in incubator at 37°C, 5%CO2 for 70±12 minutes and transplanted. Then, all Eppendorf tubes, DFCSSGs and Mp96 were washed from cells, which were counted with Hemocytometer with Trypan blue exclusion. As a result, utilisation of DFCSSG increased cellularization rate of grafts by 36.39±2.63% (p˂0.001). A difference between cells viability depending on devices for grafts cellularization (Mp96 =90.73%±2.54%, DFCSSG =89.41%±4.40%), is absent (p˃0.5).