Characterization and Optimization of the Seeding Process of Adipose Stem Cells on the Polycaprolactone Scaffolds

Abstract

The purpose of the current study was to evaluate the usefulness of adipose-derived stem cells (ASCs) for bone injury therapy. Lipoaspirates were collected from the abdomen regions of 17 healthy female donors (mean age 49 ± 6 years) using Coleman technique or Body-jet liposuction. In the present study, the primary objective was the in vitro characteristics of human ASCs. The secondary objective was the optimization of the cell seeding process on 3D-printed scaffolds using polycaprolactone (PCL) or polycaprolactone covered with tricalcium phosphate (PCL + 5% TCP). Biological evaluation of human ASC showed high efficiency of isolation obtaining a satisfying amount of homogeneous cell populations. Results suggest that ASCs can be cultured in vitro for a long time without impairing their proliferative capacity. Growth kinetics shows that the highest number of cells can be achieved in passage 5 and after the 16th passage; there is a significant decrease of cell numbers and their proliferative potential. The percentage of colony forming units from the adipose stem cells is 8% ± 0.63% (p<0.05). It was observed that the accumulation of calcium phosphate in the cells in vitro, marked with Alizarin Red S, was increased along with the next passage. Analysis of key parameters critically related to the cell seeding process shows that volume of cell suspension and propagation time greatly improve the efficiency of seeding both in PCL and PCL + 5% TCP scaffolds. The cell seeding efficiency did differ significantly between scaffold materials and cell seeding methods (p<0.001). Increased seeding efficiency was observed when using the saturation of cell suspension into scaffolds with additional incubation. Alkaline phosphatase level production in PCL + 5% TCP scaffold was better than in PCL-only scaffold. The study results can be used for the optimization of the seeding process and quantification methods determining the successful implementation of the preclinical model study in the future tissue engineering strategies.

Heterogeneity of mesenchymal stem cells in cartilage regeneration: from characterization to application

“…When rabbit ADSCs (rADSCs) or their secretome were injected into the knees of rabbits with cartilage lesions, better cartilage regeneration was achieved in the ADSC group than in the secretome group 82 , indicating that ADSCs may be more effective in the repair of cartilage damage than their secretome. In addition, many types of biomaterial scaffolds based on chitosan, fibrin, atelocollagen, and decellularized cartilage have been used as carriers to provide a 3D culture environment for ADSCs, which could support the survival, proliferation and differentiation of ADSCs 83 , 84 . For example, Boyer et al 85 seeded hADSCs on an injectable hydrogel composed of silanized hydroxypropyl cellulose to repair osteochondral defects in dogs and demonstrated satisfactory effects after 4 months.…”

Bioactivity Comparison of Electrospun PCL Mats and Liver Extracellular Matrix as Scaffolds for HepG2 Cells

“…The overall low cell attachment could be explained by the imperfection of the chosen seeding method, considering the fact that the scaffolds were not well fixed at the bottom of the wells and consequently a certain amount of the cells could slip under scaffolds after seeding. In our future experiments we will try to prevent this by anchoring the disc shaped matrices with metal rings or certain non-toxic adhesives [ 31 ]. This shortcoming of the cell seeding technique was actually the main reason why we performed all the absorbance assays in sextuplicate.…”

Characterized the Adipogenic Capacity of Adipose-Derived Stem Cell, Extracellular Matrix, and Microenvironment With Fat Components Grafting

“…Many studies had shown that without adipose-inductive scaffolds and microenvironment, transplantation of ASCs does not generate adipose tissue ( Acil et al, 2014 ; Kurzyk et al, 2019 ). However, we found that they played an important role in activating the adipoinduction potential of the ECM and microenvironment.…”