SCERG has greatly expanded the understanding of stem cell expansion and differentiation in bioreactors and pioneered the use of microcarrier technology for multipotent [1], and pluripotent stem cells [2]. Animal-free microcarrier-based stirred culture systems, envisaging xeno-free culture systems for scalable cell production, have been established for human mesenchymal stem/stromal cells [1, 3, 4, 5], and human induced pluripotent stem cells [6, 7].
- Microcarriers are coated with specific ECM molecules and peptides potentiating cell adhesion and growth in feeder-free and xeno-free conditions. This is expected to have a major impact on mimicking the stem cell niche and therefore maximizing cell growth/differentiation.
- Large-scale expansion and differentiation of stem cells in controlled microcarrier-based bioreactors are addressed through rational reaction engineering approaches for a systematic analysis of stem cells production, including kinetic studies of cell growth/metabolism, hydrodynamics/mass transfer effects on cell phenotype, oxygen tension control and feeding regimen, namely continuous perfusion of culture medium to allow a more precise control over the concentrations of soluble compounds (nutrients, toxic metabolites, growth factors). The operation of novel disposable bioreactors, with low shear stress has been optimized for the scalable xeno-free microcarrier-based cultivation of hiPSC. Innovative bioreactor designs towards the maximization of cell yield has also been performed for HSC expansion.
Selected publications:
[1] dos Santos, F., Andrade, P.Z., Abecasis, M.M., Gimble, J.M., Chase, L.G., Campbell, A.M., Boucher, S., Vemuri, M.C., da Silva, C.L., Cabral, J.M.S. Toward a clinical-grade expansion of mesenchymal stem cells from human sources: a microcarrier-based culture system under xeno-free conditions. Tissue Eng Part C Methods 17(12):1201-1210 (2011).
[2] Fernandes-Platzgummer, A., Diogo, M.M., Baptista, R.P., da Silva, C.L., Cabral, J.M.S. Scale-up of mouse embryonic stem cell expansion in stirred bioreactors. Biotechnol Prog 27(5):1421-1432 (2011).
[3] Carmelo, J.G., Fernandes-Platzgummer, A., Cabral, J.M.S., da Silva, C.L. Scalable Ex Vivo expansion of human mesenchymal stem/stromal cells in microcarrier-based stirred culture systems. Methods Mol Biol 1283, 147-159 (2015).
[4] dos Santos, F., Campbell, A., Fernandes-Platzgummer, A., Andrade, P.Z., Gimble, J.M., Wen, Y., Boucher, S., Vemuri, M.C., da Silva, C.L., Cabral, J.M. A xenogeneic-free bioreactor system for the clinical-scale expansion of human mesenchymal stem/stromal cells. Biotechnol Bioeng 111:1116-1127 (2014).
[5] Mizukami, A., Fernandes-Platzgummer, A., Carmelo, J.G., Swiech, K., Covas, D.T., Cabral, J.M.S., da Silva, C.L. Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells. Biotechnol J 11:1048-1059 (2016).
[6] Badenes, S.M., Fernandes, T.G., Cordeiro, C.S.M., Boucher, S., Kuninger, D., Vemuri, M.C., Diogo, M.M., Cabral, J.M.S. Defined essential 8™ medium and vitronectin efficiently support scalable xeno-free expansion of human induced pluripotent stem cells in stirred microcarrier culture systems. PLoS One 11:e0151264 (2016).
[7] Badenes, S.M., Fernandes, T.G., Miranda, C.C., Pusch-Klein, A., Haupt, S., Rodrigues, C.A.V., Diogo, M.M., Brüstle, O., Cabral, J.M.S. Long-term expansion of human induced pluripotent stem cells in a microcarrier-based dynamic system. J Chem Technol Biotechnol 92:492-503 (2017).