Culture of Cells: Basic Principles

Abstract

The basic techniques for cell culture are introduced in this chapter. Primarily, the requirements for cell culture are an air-conditioned room of cleanroom class 10,000 (FS209E) equipped with lights and ventilation, a laminar flow hood, a CO₂ incubator, sterilizers with both saturated steam and dry heat, a low speed centrifuge with refrigerator, upright and inverted phase contrast microscopes, a freezer at −20, a deep freezer at − 80 ̊C, a refrigerator at 4 ̊C, a sink, disposable sterile plasticware (flasks, dishes, tubes, and pipettes), balance, ultrapure water, and a supply of media, and other reagents needed for the cell environment. Further helpful apparatuses are a pH meter, a cell counter (hemocytometer), a vacuum pump, a pipette-aid, micropipettes, a liquid nitrogen tank, a fluorescent microscope, and so on. Attention to safety and the maintenance of equipments is essential to understand the significance, the reasons, and the mechanisms. Contamination of microbials such as bacteria, yeast, staphylococcus, fungus, and mycoplasma should be strictly avoided in cell culture. Simultaneously careful attention not to overgrow but grow with sufficient cell density and to avoid passages for a long time because the phenotype of the cell may subject to change. This knowledge will help researchers with even a little prior experience to set up a suitable laboratory for basic cell culture.