Colony forming cell (CFC) assay

WW Wen-Tao Wang
TC Tian-Qi Chen
ZZ Zhan-Cheng Zeng
QP Qi Pan
WH Wei Huang
CH Cai Han
KF Ke Fang
LS Lin-Yu Sun
QY Qian-Qian Yang
DW Dan Wang
XL Xue-Qun Luo
YS Yu-Meng Sun
YC Yue-Qin Chen
ask Ask a question
Favorite

A total of 10,000 MLL leukemia patient cells or CD34+ MLL leukemia cells were used per CFC assay and were added to 3 mL of methylcellulose media (R&D, USA). The vial was vigorously vortexed to thoroughly mix cells with the media. The media was allowed to stand until air bubbles disappeared. Next, we added 1.1 mL of the final cell mixture to each 35-mm culture dish. Two sample dishes and an uncovered dish containing 3–4 mL of sterile water were then placed into a 100-mm culture dish with a cover. Cells were incubated at 37 °C and 5% CO2, and the total colonies were counted after 7 days post plating.

Colony assay for MLL leukemia cell lines MOLM13 and THP1 was performed by plating 5000 cells on methylcellulose media (R&D, USA). Cells were incubated for 14 days respectively at 37 °C and 5% CO2.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A