Reproducible culture and differentiation of mouse embryonic stem cells using an automated microwell platform☆

Highlights • We describe an automated platform for hands-free ESC expansion and differentiation.• Key bioprocess variables were investigated to optimize culture inductions.• Cell growth was more consistent with automated ESC expansion than manual culture.• ESCs expanded on the automated platform maintained high levels of pluripotency.• Cells expressed βIII-tubulin after successful automated neuronal differentiation.

[1]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[2]  H. Moore,et al.  The medium is the message , 2006, Nature Biotechnology.

[3]  G. Lye,et al.  Lowering oxygen tension enhances the differentiation of mouse embryonic stem cells into neuronal cells , 2009, Biotechnology progress.

[4]  Marshall McLuhan,et al.  The medium is the message , 2005 .

[5]  Austin G Smith,et al.  Defined conditions for neural commitment and differentiation. , 2003, Methods in enzymology.

[6]  A. Smith,et al.  Embryo-derived stem cells: of mice and men. , 2001, Annual review of cell and developmental biology.

[7]  X. Chen,et al.  The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells , 2006, Nature Genetics.

[8]  Steven L. Stice,et al.  Preserving the genetic integrity of human embryonic stem cells , 2005, Nature Biotechnology.

[9]  Chris Denning,et al.  Improved genetic manipulation of human embryonic stem cells , 2008, Nature Methods.

[10]  David J. Williams,et al.  Cell Culture Automation and Quality Engineering: A Necessary Partnership to Develop Optimized Manufacturing Processes for Cell-Based Therapies , 2008 .

[11]  Chris Mason,et al.  Translational regenerative medicine research: essential to discovery and outcome. , 2007, Regenerative medicine.

[12]  S. Chandran,et al.  Automated Mechanical Passaging: A Novel and Efficient Method for Human Embryonic Stem Cell Expansion , 2006, Stem cells.

[13]  K. Byrne,et al.  Ten commandments for preventing contamination of primary cell cultures. , 2000, Methods in cell science : an official journal of the Society for In Vitro Biology.

[14]  Chris Mason,et al.  Regenerative medicine bioprocessing: building a conceptual framework based on early studies. , 2006, Tissue engineering.

[15]  F. Berthod,et al.  Differentiation of human adult skin‐derived neuronal precursors into mature neurons , 2007, Journal of cellular physiology.

[16]  Michael S Kallos,et al.  Expansion of undifferentiated murine embryonic stem cells as aggregates in suspension culture bioreactors. , 2006, Tissue engineering.

[17]  D J Williams,et al.  Application of process quality engineering techniques to improve the understanding of the in vitro processing of stem cells for therapeutic use. , 2008, Journal of biotechnology.

[18]  J. Miyazaki,et al.  Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells , 2000, Nature Genetics.

[19]  Joaquim M S Cabral,et al.  Expansion of mouse embryonic stem cells on microcarriers , 2007, Biotechnology and bioengineering.

[20]  G. Martin,et al.  Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Kowalski,et al.  Controlling Embryonic Stem Cell Growth and Differentiation by Automation , 2012, Journal of biomolecular screening.

[22]  S. Chandran,et al.  Embryonic Stem Cell‐Derived Neural Progenitors Display Temporal Restriction to Neural Patterning , 2006, Stem cells.

[23]  C. Heeschen,et al.  Genetically engineered stem cells for therapeutic gene delivery. , 2007, Current gene therapy.

[24]  H. Thomson,et al.  Bioprocessing of embryonic stem cells for drug discovery. , 2007, Trends in biotechnology.

[25]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[26]  J. Mcneish.  Embryonic stem cells in drug discovery , 2004, Nature Reviews Drug Discovery.

[27]  C. Mason,et al.  A brief definition of regenerative medicine. , 2008, Regenerative medicine.

[28]  Roy M. Williams,et al.  The ACTCellerate initiative: large-scale combinatorial cloning of novel human embryonic stem cell derivatives. , 2008, Regenerative medicine.

[29]  Richard F. Martin,et al.  Effects of stimulation in nucleus reticularis gigantocellularis on the activity of spinothalamic tract neurons in the monkey , 1978, Brain Research.

[30]  Sean P. Palecek,et al.  3-D microwell culture of human embryonic stem cells. , 2006, Biomaterials.

[31]  Wu Ma,et al.  Neural stem cell differentiation in a cell-collagen-bioreactor culture system. , 2004, Brain research. Developmental brain research.

[32]  Austin G Smith,et al.  Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture , 2003, Nature Biotechnology.

[33]  J. Ringe,et al.  A Microcarrier‐Based Cultivation System for Expansion of Primary Mesenchymal Stem Cells , 2007, Biotechnology progress.

[34]  A. Bradley,et al.  Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines , 1984, Nature.

[35]  David J. Williams,et al.  Automated, scalable culture of human embryonic stem cells in feeder‐free conditions , 2009, Biotechnology and bioengineering.

[36]  P. Andrews,et al.  OCT4 Spliced Variants Are Differentially Expressed in Human Pluripotent and Nonpluripotent Cells , 2008, Stem cells.

[37]  Chris Mason,et al.  The impact of manual processing on the expansion and directed differentiation of embryonic stem cells , 2008, Biotechnology and bioengineering.

[38]  B. Thiers.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2008 .

[39]  M. S. Levy,et al.  Framework for the Rapid Optimization of Soluble Protein Expression in Escherichia coli Combining Microscale Experiments and Statistical Experimental Design , 2007, Biotechnology progress.

[40]  Anne E. Bishop,et al.  Antibiotics reduce the growth rate and differentiation of embryonic stem cell cultures. , 2006, Tissue engineering.

[41]  Radu Dobrin,et al.  Dissecting self-renewal in stem cells with RNA interference , 2006, Nature.

[42]  Chris Mason,et al.  International community consensus standard for reporting derivation of human embryonic stem cell lines. , 2007, Regenerative medicine.

[43]  Oliver Brüstle,et al.  Automated maintenance of embryonic stem cell cultures , 2007, Biotechnology and bioengineering.

[44]  D. Kirouac,et al.  The systematic production of cells for cell therapies. , 2008, Cell stem cell.

[45]  David J. Williams,et al.  Human cell culture process capability: a comparison of manual and automated production , 2009, Journal of tissue engineering and regenerative medicine.

[46]  K. Boheler,et al.  Pluripotency of embryonic stem cells , 2007, Cell and Tissue Research.

[47]  J. Mcneish.  Stem cells as screening tools in drug discovery. , 2007, Current opinion in pharmacology.