细胞粘连问题。

爱白衣杀手

我做的组织消化细胞，发现细胞老是呈团，很粘稠，怎么办？是在FACS buffer里面加EDTA吗？什么浓度？有做过的吗？

niwanmao

从国外protocol-online网站找到相关方法，供参考。

两种解决方法：
1) EDTA only Method:

Make a solution of 1mM EDTA (can be increased all the way up to 10-15mM EDTA if your cells are extremely adherent). The EDTA solution acts to chelate calcium. Flood flask with the solution and incubate for 1 minute. Remove most of solution (not as dangerous for cells to sit in EDTA as it is to sit in trypsin) and allow to incubate at 37degrees for 5-10 minutes. Time may be increased if necessary.

I found that the EDTA solution at 10mM worked well for 10 minutes with Hep2 epithelial cells, but I had difficulty with clumping for my T84 cell line. Normally this would not be terrible, but for flow cytometry it is very problematic.

2) Citric Saline Method

For 500mL 10x solution: 50.3 g KCl, 22.06 g Sodium Citrate.

Flood flask with prewarmed citric saline (1x) and incubate at 37 degrees for 4 minutes. Remove cells be gently tapping or pipetting up and down several times.

I have not had the chance to try this yet, so please let me know how this works for you!