I want to estimate the number of tumor cells by utilizing an experimental in vivo, 4T1 cell line, tumor volume measurement. How I could do this?
The tumour was measured with a calliper and its volume was calculated using following formula: V (mm3) = (a × b 2)/2, where a and b represented maximum and minimum diameter, respectively.
Check if this paper helps you or contact the researchers, they may be on RG:
J Cell Mol Med. 2018 Dec; 22(12): 5939–5954. Published online 2018 Oct 5.
Adenosine deaminase inhibition suppresses progression of 4T1 murine breast cancer by adenosine receptor‐dependent mechanisms
Barbara Kutryb‐Zajac, 1 Patrycja Koszalka, 2 Paulina Mierzejewska, 1 Alicja Bulinska, 1 Magdalena A. Zabielska, 1 , 3 Karolina Brodzik, 2 Aleksandra Skrzypkowska, 2 Lukasz Zelazek, 2 Iwona Pelikant‐Malecka, 1 Ewa M. Slominska, 1 and Ryszard T. Smolenski📷 1
To do this with flow cytometry you could weigh/measure the tumors, and then harvest them. Next digest them into single cell suspension. Then perform flow cytometry using a panel containing at minimum (CD3, CD45, Live Dead). I use counting beads (Accucheck) for quantification per gram of tissue, where you spike in a known number of beads prior to analysis, and then based off the number you acquire you know the percentage of the cells you acquired.
For example, lets say you harvest a tumor and it weight 500mg, and after processing you have 2e6 cells to stain. You decide you only want to stain 1e6, so you stain 50% of you sample. You then stain your sample, and spike in counting beads. After analyzing the data you determine that you acquired 50% of the stained sample, and there were 5e4 T cells. Since you acquired half your sample, and stained half your tumor, you would multiply this by 4. This would tell you that there were 2e5(t cells)/.5(g) tumor. You would present this as 4e5 t cells per g of tumor, and repeat this analysis in parallel for all your tumor samples.
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