An integrated method for cell isolation and migration on a chip

Abstract

Tumour cell migration has an important impact on tumour metastasis. Magnetic manipulation is an ascendant method for guiding and patterning cells. Here, a unique miniaturized microfluidic chip integrating cell isolation and migration assay was designed to isolate and investigate cell migration. The chip was fabricated and composed of a magnet adapter, a polytetrafluoroethylene(PDMS) microfluidic chip and six magnetic rings. This device was used to isolate MCF-7 cells from MDA-MB-231-RFP cells and evaluate the effects of TGF-β on MCF-7 cells. First, the two cell types were mixed and incubated with magnetic beads modified with an anti-EpCAM antibody. Then, they were slowly introduced into the chip. MCF-7 cells bond to the magnetic beads in a ring-shaped pattern, while MDA-MB-231-RFP cells were washed away by PBS. Cell viability was examined during culturing in the micro-channel. The effects of TGF-β on MCF-7 cells were evaluated by migration distance and protein expression. The integrated method presented here is novel, low-cost and easy for performing cell isolation and migration assay. The method could be beneficial for developing microfluidic device applications for cancer metastasis research and could provide a new method for biological experimentation.

Plug-and-Play In Vitro Metastasis System toward Recapitulating the Metastatic Cascade

“…It is worth pointing out that an in vitro tumor microenvironment can be mimicked by culturing multiple cell types to generate complicated physical and biological cell-cell interactions, enabling the regulation of the invasion and metastasis of cancer cells through molecular signaling 42 . Recently, TGF-β1 has been shown to play a critical role in tumor progression and metastasis 43–45 , while a co-culture of microvascular endothelial cells is able to promote the growth of cancer cells 46 . Therefore, to quantify the actual intravasation rate of the cancer cells, the A549-GFP cells were cultured directly on the porous membrane in the U-well with a seeding density of 1.6 × 10 5 cells/mL.…”

“…S5). To verify the cellular signaling acting on the metastasis chip, 5 ng/mL (0.4 nM) of the recombinant human TGF-β1 (R&D Systems, USA) was added to the culture medium on a daily basis until the end of the experiment 45 . After seeding for 1 d, the U-wells were inserted into the metastasis chip housing, which was connected to a perfusion system to generate the dynamic and circulating flow environment.…”

Exosome Isolation and Detection: From Microfluidic Chips to Nanoplasmonic Biosensor

“…Microfluidics, where fluids are manipulated in microchannels, is a technique to proceed with biomedical and biochemical reactions . It plays an important role in disease diagnostics and therapeutics due to its unique advantages such as low cost, low sample volumes, low consumption, high precision, high throughput, real-time and point-of-care-test, and easy integration with other technologies. , Microfluidic chips can accurately control the reaction conditions at the micron level, thereby improving the reaction efficiency. The laminar flow in the microfluidic chip can provide better kinetic control for the surface modification of biosensors.…”

A Flexible Terahertz Metamaterial Biosensor for Cancer Cell Growth and Migration Detection

“…Cell migration provides important information in various biological processes. Studies have demonstrated that TGF-β could promote MDA−MB−231 cell migration [29]. To expand the application of the biosensor, the effect of TGF-β on MDA−MB−231 cells migration was investigated without any cell damage by the biosensor.…”