title: Xeno-free production of human embryonic stem cells in stirred microcarrier systems using a novel animal/human-component-free medium.
abstract: Currently, stem cell research faces a major bottleneck related to the low efficiency of methods to produce large quantities of human embryonic stem cells (ESC) for use in clinical trials. Most culture media currently employed for human ESC cultivation contain animal compounds, and cells are grown in static flasks. Besides the immediate contamination with nonhuman compounds, cell expansion in flasks tends to be laborious and nonefficient. Here we cultured human ESC in stirred microcarrier (MC) systems using an animal/human-component-free medium, to overcome both issues. The method developed to culture cells on suspended beads combined the use of polymeric MCs in stirred vessels with an optimized culture medium free of supplements of animal and human origin. This approach generated approximately 160 million cells within 6 days, which were shown to remain pluripotent. The process developed herein provides a step forward toward therapy due to the economic advantages in the production of human ESC and to their consequent low immunogenic potential.
authors: National Laboratory for Embryonic Stem Cell Research, Institute of Biomedical Sciences, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors: Paulo André Nobrega
authors: PA
authors: Marinho
authors: Daniel Tait
authors: DT
authors: Vareschini
authors: Ismael Carlos
authors: IC
authors: Gomes
authors: Bruna da Silveira
authors: Bda S
authors: Paulsen
authors: Daniel Rodrigues
authors: DR
authors: Furtado
authors: Leda dos Reis
authors: Ldos R
authors: Castilho
authors: Stevens Kastrup
authors: SK
authors: Rehen
mesh_terms: D000818:Animals; D001483:Base Sequence; D019149:Bioreactors; D002460:Cell Line; D003470:Culture Media; D017931:DNA Primers; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D008855:Microscopy, Electron, Scanning; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D003470:Culture Media; D017931:DNA Primers