Cell proliferation / Cytotoxicity Selection guide DOJINDO LABORATORIES

Targets in Cell Proliferation, Viability, and Cytotoxicity

Various assays are used for assessing cell proliferation, viability, and cytotoxicity. Examples include optical counting, [3H] thymidine, MTT, WST, lactate dehydrogenase (LDH) assay, and more. This selection guide aims to provide you with information that will allow you to pick the method most suitable for your specific experimental needs.

Proliferation

Measure cell count and/or DNA content to reveal cell division and DNA replication, respectively

Methods:

- Optical counting with hemocytometer

- DNA quantification using fluorophores

Viability

Measure parameters of cellular health to determine the intracellular impact of cytotoxicity

Methods:

-Measuring evidence of cellular metabolic activity such as esterase activity, ATP,

NADH dehydrogenase activity via MTT or WST, and so on.

Cytotoxicity

Measure indicators of cell death to evaluate cytotoxicity

Methods:

-Measure the activity of lactate dehydrogenase leaked out of damaged cell membranes

-Use membrane-impermeable dyes to selectively stain inside cells

with damaged membranes

Learn more about cell analysis

(“Reagent Selection Guide, Cell Proliferation / Cell Viability / Cytotoxicity ”)

Chapters:

0:00 Title
1:11 What Are Cell Proliferation, Viability and Cytotoxicity Tests?
4:12 Cell Viability: Comparison of Each Reagent and Assay Principle
8:53 Cytotoxicity: Comparison of Each Reagent and Assay Principle
13:52 Measurement of Multiple Parameters
16:33 More Detailed Cellular Function Analysis

Cell Proliferation / Cytotoxicity Test – Related Reagents

The MTT and WST techniques are widely-used methods for evaluating cell proliferation and cytotoxicity. They are valued for the simplicity of their respective procedures, their safety, and the reproducibility of their results. WST (water-soluble tetrazolium) was developed by DOJINDO Laboratories in the 1990's and is now used worldwide in cell proliferation assays. Cell Counting Kit-8 (CCK-8) is a commercial product that utilizes the WST method with improved sensitivity. Cell Counting Kit-8 is the original WST product, with which more than 5,000 papers have been published worldwide.

Product name	MTT	Cell Counting Kit-8	ATP Assay Kit	Cell Counting Kit-F Calcein-AM
Target	Mitochondrial dehydrogenase activity	Cellular dehydrogenase activity	ATP	Esterase activity
Instrument	Microplate reader (colorimetric)	Microplate reader (colorimetric)	Microplate reader (luminometric)	Microplate reader, FCM, Fluorescence microscope (fluorometric)
Advantage	Less expensive	Simple procedure High stable reagent	High sensitivity Short reaction time(15 min) Simple procedure	Applicable for various instruments Short reaction time(15 min)
Disadvantage	Affected by reducing agents Requires dissolution of reagent after reaction	Affected by　reducing agents	Expensive	Serum increases background Leakage of reagent
Product code	M009	CK04	-	CK06 (kit) C396 (ready-to-use solution) C326 (powder)

Please watch this video for comparison between MTT and Cell Counting Kit-8.

Cell Proliferation Cytotoxicity Assay Kit_CCK-8

Cell Counting Kit-8 VS MTT

Comparison of Each Cytotoxicity Assay Reagent

Cytotoxicity LDH Assay Kit-WST is a kit used for evaluating cytotoxicity. The assay measures lactate dehydrogenase released from damaged cell membranes. Cytotoxicity LDH Assay Kit-WST is well-suited for multi-sample assays, and the associated procedure for evaluating cytotoxicity is simple.

Product name	Cytotoxicity LDH Assay Kit-WST	Cell Counting Kit-F (Calcein-AM)	Propidium iodide (PI)	Trypan Blue
Target	Released Lacate dehydrogenase (LDH)	A marker (Calcein-AM) released into the supernatant (Calcein-Release Assay)	Cytoplasm(Live cell membranes are impermeable to PI, but not dead cell membranes)	Cytoplasm; cells taking up trypan blue are considered non-viable
Instrument	Microplate Reader (colorimetirc)	Microplate reader, FCM, Fluorescence microscope (Fluorometric)	Microplateplate reader, FCM, (fluorometric)	Microscope (colorimetric)
Advantage	・Simple procedure ・Suitable for multi-sample assay	・Convenient for cytotoxicity assay using 2 kinds of cell samples	・Less expensive ・Simple procedure	・Less expensive ・No special instruments needed
Disadvantage	・Increased background by serum	・Leakage of reagent from living cells	・Not applicable for multi-samlple assay	・False-positive by cytotoxicity ・Not applicable for multi-samples assay
Product Code	CK12	CK06 (kit) C396 (ready-to-use solution) C326 (powder)	P378 (ready-to-use solution) P346 (powder)	-

Measurement with multiple parameters in cytotoxicity assay

In order to verify the accuracy of results in a cytotoxicity assay, individual samples are measured according to multiple different principles. Cell Counting Kit-8 (code: CK04) and Cytotoxicity Assay Kit-WST (code: CK12) measure dehydrogenase activity and released amount of LDH as reflective of living and dead cells, respectively. According to these results, the number of living cells was reduced, and the number of dead cells increased as the concentration of toxicant increased.