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Cell Culture Guide

Do you need help with cell culture?


Read on to learn about choosing the optimal cell culture media for different types of cells in your laboratory. Before we introduce the mostly used types of culture media, let us start with defining cell culture and why it is required.


Cell culture dish
Cell culture dish

What is cell culture?


Cell culture is a process of regrowing cells taken from living organisms' tissues . Plant callus, a tissue involved in plant healing, is commonly cultured as are animal tumor cell lines. The reason why tumor cells are preferred and more useful is due to their accelerated growth rate and low mortality rate compared to other cells.


The purpose of cell culture is to:


1. Produce proteins for therapy purposes, including enzymes, hormone vaccines, anti-cancer drugs, and immunological regulatory factors.


2. To grow organs or tissues in vitro.


Cell culturing process


1. Thawing cells: The cells in the nitrogen tank can be thawed by submerging them in a 37°C water bath for approximately 60 seconds.


2. Detaching cells: Chemical procedures, rather than physical scraping, are used to remove cells from the bottom of a culture dish while causing minimal damage. In this technique, trypsin and EDTA are used.


3. Counting cells: Cell counting measures cell viability. This method uses either the classic hemocytometer or automated cell counters.


To understand the distinction between manual and automated cell counting, click


Manual counting:


Auto counting:


4. Incubating cells: After transferring the cells to a new cell culture dish, incubate them at 37 °C.


Types of cell culture media


The materials needed for cells to grow and survive are provided by cell culture medium. For best results, different culture media should be utilized for different cell types.


Commonly used cell culture media are:

1.     Basal medium – a mixture of amino acids, glucose, and ions


A.    Dulbecco’s Modified Eagle Medium (DMEM) : a basal medium widely used for adherent cell lines. This medium contains glutamine, glucose, sodium pyruvate and sodium bicarbonate. This is used for cell culturing and to observe cell growth and viability [1]. Cells that require DMEM for culturing or growth are cells lines such as HeLa, HEK293, Cos-7 and PC-12 [2].


B.    RPMI 1640 : a medium specifically used for the growth of suspension cells, particularly lymphocytes which contains glutathione and vitamins that are not included in other media. Cells that use this media are mammalian cells such as HeLa, Jurkat, MCF-7, PC-12 and PBMC [3].


C.    Minimum Essential Medium (MEM) : a medium similar to DMEM but contains higher concentrations of components. It can support the growth of a wide range of mammalian cells such as HeLa, HEP-2, MCF-7 and fibroblast [4].


D.    Ham’s F-12 : a medium specifically made for single-cell plating of Chinese Hamster Ovary (CHO) cells [5].


2.     Media with animal serum

1.     Fetal Bovine Serum (FBS) : a medium mostly used for cell culturing which is derived from the blood of cow fetus [6].


Most media include L-glutamine which is an amino acid necessary for protein and nucleic acid synthesis and energy production in cell culture [7].




References:

1.Dulbecco’s Modified Eagle’s Medium (DMEM) - 30-2002 | ATCC, www.atcc.org/products/30-2002. Accessed 26 Mar. 2024.

4.“Minimum Essential Medium (MEM).” Thermo Fisher Scientific - US, www.thermofisher.com/kr/ko/home/life-science/cell-culture/mammalian-cell-culture/cell-culture-media/mem.html. Accessed 26 Mar. 2024.

5.“Ham’s F-12 Nutrient Mix.” Thermo Fisher Scientific - US, www.thermofisher.com/order/catalog/product/kr/en/11765054. Accessed 26 Mar. 2024.

7.Author links open overlay panelJessica J. Alm 1, et al. “Clinical Grade Production of Mesenchymal Stromal Cells.” Tissue Engineering (Second Edition), Academic Press, 23 Jan. 2015, www.sciencedirect.com/science/article/abs/pii/B9780124201453000134?via%3Dihub.





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