We are developing a technique that can isolate circulating tumor cells (CTCs) from patient blood. I wonder if we can culture isolated CTC cells starting from very few cells since normally there are less than 100 CTCs in 10 ml. of blood.
Dear Sittisak
The number of the isolated CTCs is 1-10 / ml of whole blood. The problem is the
VIABILITY of these cells.
I found a publication for you:
"Rapid isolation of viable circulating tumor cells from patient blood samples," by Hughes A.D. et al. J. Vis. Exp. (64) e4248 doi 10379L 4248 (2012).
I hope it can be helpful to you.
with best wishes
Eva
Thank you Eva for your suggestion.
As said in paper "Rapid isolation of viable CTCs..." likely that I am using the same culture media, RPMI1640+20%FBS+antibiotics. However, as you stated the number of isolsted CTCs in my case was very fews which may cause CTCs to die very soon. Also contamination with microorganisms is my major concern here.
Dear Sittisak
I wrote you that the viability of the CTCs is impaired. These cells die very quickly in the blood and in the culture medium. The CTCs are so-called semi-dead cells.
My question: What type of cancer and in which stage(s) have the patients? I could have an idea....
Best
Eva
Dear EVa
We have been trying to isolate CTCs from cholangiocarcinoma patient bloods. Either blood from M0 or M1 stage were tested but as far as we go is that some attached cells look like macrophages grown and then die within 1 month. You may watch its picture from this topic https://www.researchgate.net/post/What_type_of_cell_is_it_image_attached?ch=reg&cp=re72_x_p2&pli=1&login=sittisak@cri.or.th#view=50e55273e5438fd31200000c
Our possible concerns is that developing separation technique is not good enought or few isolated CTCs could not be grown in RPMI+20%FBS media. Likely there are several issues need to be taken into account.
Dear Sittisak
To my histological knowledge this is an endothelial cell. It seems, this is a dead cell...
I think your problem is not a technical problem. The problem lies in the tumor cells. These tumor cells come down generally by passive shedding. These circulating tumor cells are practically dead. It is imaginable that aggressive and poorly differentiated or undifferentiated tumors have a directed migration mechanism, leading to aggressive and viable shedding-cells. Ask the pathologist!
Second possibility; the culture medium. The DSMZ cultivate tumor cells to complete commercial tumor cell lines. About 100 passages are necessary. It is possible they use an other medium than RPMI.....
You can contact in this connection: Steube Klaus, MD, email: [email protected] He is very kind and helpful.
Have a nice weekend
with best wishes
Eva
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