A microfluidic live cell assay to study anthrax toxin induced cell lethality assisted by conditioned medium.
It is technically challenging to investigate the function of secreted protein in real time by supply of conditioned medium that contains secreted protein of interest. The internalization of anthrax toxin is facilitated by a secreted protein Dickkopf-1 (DKK1) and its receptor, and eventually leads to cell lethality. To monitor the dynamic interplay between these components in live cells, we use an integrated microfluidic device to perform the cell viability assays with real-time controlled culture microenvironment in parallel. Conditioned medium, which contains the secreted proteins from specific cell lines, can be continuously pumped towards the cells that exposed to toxin. The exogenous DKK1 secreted from distant cells is able to rescue the sensitivity to toxin for those DKK1-knocked-down cells. This high-throughput assay allows us to precisely quantify the dynamic interaction between key components that cause cell death, and provide independent evidence of the function of DKK1 in the complex process of anthrax toxin internalization.
1] Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China [2] College of Engineering, Peking University, Beijing, 100871, China [3] School of Life Sciences, Peking University, Beijing, 100871, China.
Jie
J
Shen
School of Life Sciences, Peking University, Beijing, 100871, China.
Changzu
C
Cai
1] Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China [2] College of Engineering, Peking University, Beijing, 100871, China.
Zhilong
Z
Yu
1] Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China [2] School of Life Sciences, Peking University, Beijing, 100871, China.
Yuhong
Y
Pang
1] Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China [2] College of Engineering, Peking University, Beijing, 100871, China.
Ying
Y
Zhou
School of Life Sciences, Peking University, Beijing, 100871, China.
Lili
L
Qian
School of Life Sciences, Peking University, Beijing, 100871, China.
Wensheng
W
Wei
1] Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China [2] College of Engineering, Peking University, Beijing, 100871, China.
Yanyi
Y
Huang
D000818:Animals; D000942:Antigens, Bacterial / Q000633:toxicity*; D001427:Bacterial Toxins / Q000633:toxicity*; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects* / Q000235:genetics; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D057166:High-Throughput Screening Assays; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D051379:Mice; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D034622:RNA Interference
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D017077:Culture Media, Conditioned; C501127:Dkk1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; C030325:anthrax toxin