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Change media to fresh complete DMEM (day 3). 3. 3. 6 × 106 PLAT-E cells each (one for each pMX plasmid) and incubate overnight in complete DMEM media without puromycin or blasticidin S in a standard tissue culture incubator. 3 mL of OPTI-MEM I media into three microcentrifuge tubes, add 27 mL of Fugene 6 transfection reagent to each, mix by finger tapping, and incubate for 5 min at room temperature. Add 9 mg of either OCT3/4, KLF4, or SOX2 pMX plasmid to the three microcentrifuge tubes, mix by finger tapping, and incubate 15 min at room temperature.
Altern Lab Anim 26:763–777 11. Jones BPC (1998) Laboratory practice. In: Stacey G, Doyle A, Hambleton P (eds) Safety cell and tissue culture. Kluwer Academic, Dordrecht, The Netherlands, pp 64–86 Chapter 2 Induced Pluripotent Stem Cells (iPSCs): An Emerging Model System for the Study of Human Neurotoxicology M. Diana Neely, Andrew M. Tidball, Asad A. Aboud, Kevin C. Ess, and Aaron B. Bowman Abstract This chapter describes the materials and methods necessary to generate human induced pluripotent stem cells (iPSCs) from primary human fibroblasts and direct their differentiation into neural progenitor cells.
Risk assessment is not only limited to the laboratory and laboratory personnel, but also to people in the entire facility, people in the external environment and to the environment itself. This is not only a vital aspect of basic research and testing, but is particularly important when cultured cells and tissues are used for diagnostic purposes or for producing therapeutic products, or when the cells and tissues themselves are used for therapeutic purposes. 1. Physical Hazards The cell and tissue culture laboratory does not pose any specific physical hazards.