Autoria: DUARTE, F. B.; BRÍGIDO, M. de M.; MELO, E. de O.; BÁO, S. N.; MARTINS, C. F.

Resumo: Cells from different origins behave differently regarding the incorporation of exogenous DNA and formation of transgenic cells. Milk production of recombinant antibody may benefit from efficient transfection protocols to produce transgenic animals. In this context, the objective of this study was to verify the transfection potential of bovine mesenchymal stemcells from Wharton’s jelly (MSC-WJ) and adipose tissue (MSC-AT), comparing co-transfection protocols with vectors pBC1-anti-CD3 and pEF-NEO-GFP, using transfection reagents Lipofectamine LTX with Plus Reagent or Xfect. Skin fibroblasts (FIB) were used as the control group. Forty-eight hours after transfection, neomycin was added and cells cultured for 2 weeks. Treated cells were submitted to fluorescence microscopy, flow cytometry, and PCR evaluations. Wharton’s jelly cells were sensitive to treatments and started necrosis. In theflow cytometry assay, the median fluorescence was higher in adipocytes than fibroblasts, forboth the Xfect (20.057 ± 1.620,7 and 10.601 ± 702,86, respectively, p < 0.05) and LTX(19.590 ± 113,84 and 10.518 ± 442,65, respectively, p < 0.05). These results, associated withevaluation of epifluorescence, demonstrated that adipocytes presented a better response to transfection than other cells, independent of the kit used. Performing PCR on co-transfectedcells demonstrated the presence of anti-CD3, making this approach feasible for future experiments.

Ano de publicação: 2022

Tipo de publicação: Artigo de periódico

Unidade: Embrapa Recursos Genéticos e Biotecnologia

Palavras-chave: Humanized anti-CD3, Monoclonal antibody, PBC1, Transfection