Three-Dimensional Spheroid Invasion Assay

In experimental cellular biology, invasion is defined as disruptive, proteolytic,

cellular movement through three-dimensional barriers. Despite intense study,
the complex and heterogenous mechanisms of invasion and metastasis
remain imperfectly understood and several mechanisms exist in collective or
individual invasion of cancer cells. Thus, there is a need to establish robust,
reproducible assays to target the process and get a detailed analysis of
invasion. The three-dimensional (3D) spheroid invasion assay is a simple and
convenient method to research cell invasion, which is based on the uniform
and self-assembling 3D tumor spheroids.
The three-dimensional (3D) spheroid invasion assay can provide a rapid,
automatable invasion system to determine cell invasion such as tumor cell.
This method is also highly reproducible and has many other advantages than
other invasion assays. During the process of three-dimensional (3D) spheroid
invasion assay, spheroids will be formatted in a 96-well plate and each well
will format one spheroid. A semi-solid matrix is generated by the BMM added
directly to each well into which tumor cells invade from the spheroid body.
Then the cell invasion is monitored by the imaging cytometer, which scans a
96-well plate within 10 minutes. The extent and rate of the cell invasion can
be measured in a dynamic fashion by using the confluence application,
whatever the invasion is achieved by single cell or by cell clusters spreading
out from the tumor spheroids and invading into the matrix. The determination
of cell invasion in this method is helpful for the further understanding of the
mechanisms of metastasis of cancer.
This method to study cell invasion can provide great advantages over other in
vitro cell invasion assays. In three-dimensional (3D) spheroid invasion assay,
the tumor cells are organized into a 3D culture, which could mimic a tumor
micro-region or micro-metastasis. Another advantage of this assay is high
reproducibility, since the spheroids are highly reproducible in size and this
assay is performed in the same plate without the need to move the spheroids
to another plate. In our laboratory, the application of the latest technologies of
automated image analysis enables the high throughput analysis of cell
invasion.