Table of Contents
What is the purpose of a cell migration assay?
The transwell cell migration assay measures the chemotactic capability of cells toward a chemo-attractant. The transwell cell invasion assay, however, measures both cell chemotaxis and the invasion of cells through extracellular matrix, a process that is commonly found in cancer metastasis or embryonic development.
How do you analyze cell migration?
Thus to analyze cell migration more accurately a migration assay is recommended allowing to measure the migration of single cells within a 3D physiological or physiological-like environment. Intravital imaging/microscopy is the gold-standard for measuring cell migration within a 3D physiological context.
How do you quantify migration assay?
Migration can be assessed by determining the number of cells that move across a microporous membrane (transwell migration assay) or by measuring the surface area that cells occupy over time after creating a ‘cell-free’ area (scratch assay) [8–10].
What is cell invasion assay?
These 96-well microplate-based assays are designed to accelerate the screening process for compounds that influence chemotaxis. The Cell Migration Assay measures the number of cells traversing a porous membrane, while the Cell Invasion Assays monitor cell movement through extracellular matrices.
What are two types of cell migration?
Roughly speaking, cell migration can be categorized into single-cell migration and collective cell migration. Each migration mode is then further sub-categorized into several different types of migration (Figure 1). Next to migration, cells can also display invasion.
How is cell proliferation measured?
A tried-and-true method of measuring proliferation by metabolism is using tetrazolium salts such as MTT, MTS or XTT. The salts are reduced by metabolically active cells to a colored formazan, which is then detected using a spectrophotometer.
What is MTS for cell viability?
The MTS assay is used to assess cell proliferation, cell viability and cytotoxicity. The MTS assay protocol is based on the reduction of the MTS tetrazolium compound by viable mammalian cells (and cells from other species) to generate a colored formazan dye that is soluble in cell culture media.
How do HUVECs differentiate during the assay?
During the assay, HUVEC cells differentiate, directionally migrate to align, branch, and form the polygonal networks of blood vessels. Figure 1. Effect of cell number on HUVECs tube formation. Prepare the HUVECs medium 200PRF supplement with LSGS according to the product insert.
What is the suitable transwell for HUVEC migration and invasion assay?
Secondly what should be the fixation method and staining time to stain well migrated cells The 8 micron pore size transwell is suitable for the HUVEC migration and invasion assay. The optimum seeding density and incubation time is also important. Are you pre-starving the HUVEC cells before experiment and what chemoattractant are you using in study?
How to do collective cell migration assay with umbilical vein cells?
In the experiments, human umbilical vein cells (HUVECs) are exploited for the collective cell migration assays using the microfluidic devices. Collective cell migration assays with combinations of three different oxygen conditions: normoxia, oxygen gradient, and uniform hypoxia (1%), and three medium compositions are conducted in the experiments.
Why do HUVECs move slower in cytochalasin-D?
Therefore, the slower collective cell migration speed of the cells cultured in the growth medium and cytochalasin-D in the experiments are mainly caused by the lower motility of the HUVECs. In comparison, the cell numbers are all lower than the control one for the HUVECs cultured in the HIF inhibitor, YC-1.