Description and characterization of a chamber for viewing and quantifying cancer cell chemotaxis.
Direct observations of cancer cell invasion underscore the importance of chemotaxis in invasion and metastasis. Yet, there is to date, no established method for real-time imaging of cancer chemotaxis towards factors clinically correlated with metastasis. A chamber has been designed and tested, called the Soon chamber, which allows the direct observation and quantification of cancer cell chemotaxis. The premise for the design of the Soon chamber is the incorporation of a dam, which creates a steep gradient while retaining stability associated with a pressure-driven system. The design is based on the characteristics of cancer cell motility such as relatively low speeds, and slower motility responses to stimuli compared to classical amoeboid cells like neutrophils and Dictyostelium. We tested MTLn3 breast carcinoma cells in the Soon chamber in the presence of an EGF gradient, obtaining hour-long time-lapses of chemotaxis. MTLn3 cells migrated further, more linearly, and at greater speeds within an EGF gradient compared to buffer controls. Computation of the degree of orientation towards the EGF/buffer source showed that MTLn3 cells were significantly more directional toward the EGF gradient compared to buffer controls. Analysis of the time-lapse data obtained during chemotaxis demonstrated that two populations of cancer cells were present. One population exhibited oscillations in directionality occurring at average intervals of 12 min while the second population exhibited sustained high levels of directionality toward the source of EGF. This result suggests that polarized cancer cells can avoid the need for oscillatory path corrections during chemotaxis.
Soon
Lilian
L
Albert Einstein College of Medicine, Jack and Pearl Resnick Campus, Bronx, New York, USA. lilian.soon@emu.usyd.edu.au
Mouneimne
Ghassan
G
Segall
Jeffrey
J
Wyckoff
Jeffrey
J
Condeelis
John
J
D001943:Breast Neoplasms / Q000503:physiopathology; D002277:Carcinoma / Q000503:physiopathology; D002633:Chemotaxis / Q000502:physiology; D015194:Diffusion Chambers, Culture / Q000295:instrumentation*; D006801:Humans; D009361:Neoplasm Invasiveness; D014407:Tumor Cells, Cultured