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Water for Cell Culture | what type of water should be used for cell culture?

Oct. 21, 2024
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Water is extensively used in the key procedures of cell culture process. It is mainly used as a solvent in the preparation of buffers and media, other roles including the dissolution of additives and drugs, rinsing of bioreactors, plastic ware and glassware, etc. Contaminants in water may influence the growth, morphology as well as behavior of cultured cells, thereby affecting the outcome. It is necessary to choose the right water with quality up to standards to perform cell culture. Besides, monitoring and maintaining water quality throughout the process plays an important part in running predictable and repeatable experiments. Thus, how to select a reliable water purification system for this application is one of the key concerns of researchers.

 

Effects of common impurities in tap water on cell culture

 

Impurities from tap water

Effects on cell culture

(Inorganic) Ions

Toxic to different cells

Inhibits the reactions

Cause side reactions

Bacteria

Cause pH changes in media

Contaminate incubator or hood

Endotoxins

Change cell shapes

Cell adhesion or division

Change the growth and functions of   cells

Affect cloning efficiency, and production of recombinant proteins

TOC

Affect the development of cells

 

Referring to the guidelines provided by ISO3696, CLSI (Clinical and Laboratory Standards Institute), ASTM International (American Society for Testing and Materials), EMA (European Medicines Agency) and USP (US Pharmacopeia), fresh ultrapure water (Type I water) is strongly recommended for cell cultivation, and the highest specification is required to avoid contaminants introduction, as summarized in the table below.

 

Cell Culture Grade Water requirements

Parameter

Standard

Microorganisms

< 1 cfu/100ml

Pyrogens (endotoxins)

< 0.03 EU/ml

TOC level

< 5 ppb

Resistivity   (@ 25 ℃)

≥ 18 MΩ ·   cm

 

Special care needs to be taken to make sure that the water used for cell culture is free from any inorganic ions (especially heavy metals), organic compounds, pyrogens and bacterial.




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