The robust and scalable cell manufacturing towards the cost-effective delivery of safe and potent cell-based products (either autologous or allogeneic) relies on process engineering tools to understand the impact of cellular features (biological, biochemical, etc) on cell product function and performance, and how do process variables influence the critical quality attributes of the cell product. At SCERG we are developing the innovative manufacturing of two ex-vivo expanded cell-based products:
- umbilical cord blood (UCB)-derived hematopoietic stem/progenitor cells for potential application in hemato-oncological settings. Current research is focused on (i) the definition of optimal culture conditions namely concerning cytokine combinations supplementing xeno(geneic)-/serum-free culture media [1], enrichment procedures and initial cell concentrations [2], as well as oxygen tension [3]; and (ii) the understanding of the mechanisms underlying the hematopoietic supportive capacity of human mesenchymal stem/stromal cells in a co-culture setting [4].
- mesenchymal stem/stromal cells (MSC) from different tissue sources for immunomodulation-based therapies. Human MSC from adult bone marrow (BM), adipose tissue (AT), umbilical cord matrix (UCM) and synovial membrane (SM) are studied. Proliferation kinetics and metabolism of human MSC cultured ex-vivo under different oxygen tensions have been thoroughly studied along consecutive passaging [5, 6]. Culture protocols are being optimized for the efficient isolation and expansion of MSC from the different sources [5,7,8], namely focusing the use of xeno-/serum-free culture conditions [8, 9]. In addition, a proteomic analysis platform established in collaboration with BSRG [10] is being employed to understand how the ex-vivo culture process affects MSC features at the proteome level.
Selected publications:
[1] Andrade, P.Z., dos Santos, F., Almeida-Porada, G., da Silva, C.L., Cabral, J.M.S. Systematic delineation of optimal cytokine concentrations to expand hematopoietic stem/progenitor cells in co-culture with mesenchymal stem cells. Mol BioSyst 6(7):1207-1215 (2010).
[2] Andrade, P.Z., da Silva, C.L., dos Santos, F., Almeida-Porada, G., Cabral, J.M.S. Initial CD34+ cell-enrichment of cord blood determines hematopoietic stem/progenitor cell yield upon ex-vivo expansion. J Cell Biochem 112(7):1822-1831 (2011).
[3] Andrade, P.Z., de Soure, A.M., dos Santos, F., Paiva, A., Cabral, J.M.S, da Silva, C.L. Ex vivo expansion of cord blood haematopoietic stem/progenitor cells under physiological oxygen tensions: clear-cut effects on cell proliferation, differentiation and metabolism. J Tissue Eng Regen Med 9:1172-1181 (2015).
[4] da Silva, C.L., Gonçalves, R., dos Santos, F., Andrade, P.Z., Almeida-Porada, G., Cabral, J.M.S. Dynamic cell-cell interactions between cord blood hematopoietic progenitors and the cellular niche are essential for the expansion of CD34(+), CD34(+)CD38(-) and early lymphoid CD7(+) cells. J Tissue Eng Regen Med 4(2):149-158 (2010).
[5] dos Santos, F., Andrade, P.Z., Boura, J.S., Abecasis, M.M.A., da Silva, C.L., Cabral, J.M.S. Ex Vivo Expansion of Human Mesenchymal Stem Cells: A More Effective Cell Proliferation Kinetics and Metabolism Under Hypoxia J Cell Physiol 223(1):27-35 (2010).
[6] Oliveira, P.H., Boura, J., Abecasis, M.A., Gimble, J.M., da Silva, C.L., Cabral, J.M.S. Impact of hypoxia and long-term cultivation on the genomic stability and mitochondrial performance of ex vivo expanded human stem/stromal cells. Stem Cell Res 9:225-236 (2012).
[7] Santhagunam, A., Santos, F., Madeira, C., Salgueiro, J., Cabral, J. Isolation and ex-vivo expansion of Synovial Mesenchymal Stromal Cells for Cartilage Repair Cytotherapy 16(4):440-453 (2014).
[8] Simões, I.N., Boura, J.S., dos Santos, F., Andrade, P.Z., Cardoso, C.M., Gimble, J.M., da Silva, C.L., Cabral, J.M. Human mesenchymal stem cells from the umbilical cord matrix: successful isolation and ex-vivo expansion using serum-/xeno-free culture media. Biotechnol J 8(4):448-458 (2013).
[9] de Soure, A.M., Fernandes-Platzgummer, A., Moreira, F., Lilaia, C., S.-H., Ku, C.-P., Huang, Y.-F., Milligan, W., Cabral, J.M.S., Silva, C.L. Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells J Tissue Eng Regen Med 11:1630-1640 (2016).
[10] Madeira, A., da Silva, C.L., dos Santos, F., Camafeita, E., Cabral, J.M., Sá-Correia, I. Human Mesenchymal Stem Cell Expression Program upon Extended Ex-vivo Cultivation, as Revealed by 2-DE-Based Quantitative Proteomics. PLoS One 7(8):e43523 (2012).