(P 327) Quantitative Assay of Stem Cells Loaded on Porous Scaffolds using Bioluminescence

• Published in: Tissue engineering. Part A Vol. 14; no. 5; p. 901
• Main Authors: Logeart-Avramoglou, D, Oudina, K, Delpierre, L, Bour-Guignon, M, Nicola, MA, Petite, H
• Format: Journal Article
• Language: English
• Published: 01.05.2008
• ISSN: 1937-3341

In stem cell-based tissue engineering strategies, the capacity to monitor in vitro and in vivo proliferation of cells seeded on porous scaffolds is of great importance to scaffold design. We used bio-luminescence technology for quantitative cell assay inside scaffolds using cells tagged with the luciferase gene reporter. A clonal cell line exhibiting high luciferase expression was selected from murine mesenchymal stem cells genetically modified using lentiviral vectors encoding luciferase. Two materials were tested: AN69 hydrogel and porous coral scaffold. In vitro quantitative evaluation of luminescence from cells loaded scaffolds was performed in cellular destructive (measurement of light with a luminometer after cell lysis) and non-destructive (using a bio-luminescence imaging system) conditions. In vivo quantitative evaluation of cells adhered onto materials was accomplished in non-destructive conditions after subcutaneous implantation of implants in mice. In cellular destructive conditions, light emission from either cells or cells adhered onto scaffolds indicated a linear relationship with the cell number, despite attenuation of the transmitted light photons due to the presence of the material. Based on the ratio of slope values of standards plots, coefficient of transmission particular to each material scaffold was calculated as 0.27 plus or minus 0.05 for AN69 hydrogels and 0.35 plus or minus 0.03 for coral. In non-destructive conditions both in vitro and in vivo, analysis of luminescent images also provided evidence of correlation between the emitted photon flux and the number of cells adhering onto the materials. Bioluminescent imaging enabled non-invasive monitoring of the fate of cells present in each scaffold in living mice for 28 days post-implantation.