Video: Cell Culture Basics
 

Video 1:  Introduction to cell culture

This video provides an overview of the basic equipment used in cell culture and proper laboratory set-up.  Guidance on how to work safely and aseptically in a cell culture hood is introduced and demonstrated.

 Video: Cell Culture Basics
 

Video 1:  Introduction to cell culture

This video provides an overview of the basic equipment used in cell culture and proper laboratory set-up.  Guidance on how to work safely and aseptically in a cell culture hood is introduced and demonstrated.

 

Incubators

The purpose of the incubator is to provide the appropriate environment for cell growth.  The incubator should be large enough for your laboratory needs, have forced-air circulation, and should have temperature control to within ±0.2°C.  Stainless steel incubators allow easy cleaning and provide corrosion protection, especially if humid air is required for incubation.  Although the requirement for aseptic conditions in a cell culture incubator is not as stringent as that in a cell culture hood, frequent cleaning of the incubator is essential to avoid contamination of cell cultures.

Types of Incubators


There are two basic types of incubators, dry incubators and humid CO2 incubators.  Dry incubators are more economical, but require the cell cultures to be incubated in sealed flasks to prevent evaporation.  Placing a water dish in a dry incubator can provide some humidity, but they do not allow precise control of atmospheric conditions in the incubator.   Humid CO2 incubators are more expensive, but allow superior control of culture conditions.  They can be used to incubate cells cultured in Petri dishes or multiwell plates, which require a controlled atmosphere of high humidity and increased CO2 tension.

Storage

A cell culture laboratory should have storage areas for liquids such as media and reagents, for chemicals such as drugs and antibiotics, for consumables such as disposable pipettes, culture vessels, and gloves, for glassware such as media bottles and glass pipettes, for specialized equipment, and for tissues and cells.  Glassware, plastics, and specilized equipment can be stored at ambient temperature on shelves and in drawers; however, it is important to store all media, reagents, and chemicals according to the instructions on the label.

Some media, reagents, and chemicals are sensitive to light; while their normal laboratory use under lighted conditions is tolerated, they should be stored in the dark or wrapped in aluminum foil when not in use.

Refrigerators

For small cell culture laboratories, a domestic refrigerator (preferably one without an autodefrost freezer) is an adequate and inexpensive piece of equipment for storing reagents and media at 2–8°C.  For larger laboratories, a cold room restricted to cell culture is more appropriate.  Make sure that the refrigerator or the cold room is cleaned regularly to avoid contamination.
 

Freezers

Most cell culture reagents can be stored at –5°C to –20°C; therefore an ultradeep freezer (i.e., a –80°C freezer) is optional for storing most reagents.  A domestic freezer is a cheaper alternative to a laboratory freezer.  While most reagents can withstand temperature oscillations in an autodefrost (i.e., self-thawing) freezer, some reagents such as antibiotics and enzymes should be stored in a freezer that does not autodefrost.

Cryogenic Storage

Cell lines in continuous culture are likely to suffer from genetic instability as their passage number increases; therefore, it is essential to prepare working stocks of the cells and preserve them in cryogenic storage (for more information, see Freezing Cells).  Do not store cells in –20°C or –80°C freezers, because their viability quickly decreases when they are stored at these temperatures.  

There are two main types of liquid-nitrogen storage systems, vapor phase and liquid phase, which come as wide-necked or narrow-necked storage containers.  Vapor phase systems minimize the risk of explosion with cryostorage tubes, and are required for storing biohazardous materials, while the liquid phase systems usually have longer static holding times, and are therefore more economical.

Narrow-necked containers have a slower nitrogen evaporation rate and are more economical, but wide-necked containers allow easier access and have a larger storage capacity. 

Cell Counter

A cell counter is essential for quantitative growth kinetics, and a great advantage when more than two or three cell lines are cultured in the laboratory.  



The Countess® Automated Cell Counter is a benchtop instrument designed to measure cell count and viability (live, dead, and total cells) accurately and precisely in less than a minute per sample, using the standard Trypan Blue uptake technique.  Using the same amount of sample that you currently use with the hemocytometer, the Countess® Automated Cell Counter takes less than a minute per sample for a typical cell count and is compatible with a wide variety of eukaryotic cells.