细胞毒性测定方法-康奈尔大学刘瑞海实验室实验方法

Cytotoxicity Protocol

Cytotoxicity was measured using the method of Oliver et al, (1989) with modifications by our laboratory.

1.HepG2 cells are seeded at 4 × 104/well on a 96-well plate in 100 μL of Complete

Medium.

2.Incubate for 24 h at 37 °C.

3.Remove the medium, wash cells with 100 μL sterile cold PBS once.

4.Add treatments of fruit extracts or antioxidant compounds in 100 μL of Complete

Medium.

5.Incubate the plates at 37 °C for 24 h.

6.Perform Methylene Blue Counting.

7.Concentrations of pure compounds or fruit extracts that decrease the absorbance

by >10% when compared to the control are considered to be cytotoxic.

Sample: Add 100 μL apple extract with different concentrations + 100 μL Cell (including growth medium)

Control wells: Add 100 μL H2O + 100 μL Cell (including complete medium)

Blank wells: Add 100 μL growth medium (without cell) + 100 μL H2O (种板时就加上还是？)

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