Biocompare Bench Tip – 3D Cell Culture

Biocompare Bench Tip – 3D Cell Culture

Over the last decade a central focus for
improving drug efficacy in clinical trials has been to increase the
biological relevance of assays performed early in the drug discovery process.
However it remains difficult to simulate a drug in vivo response using in in
vitro assay. Today we’re going to focus on the importance of 3D cell culture and
the impact it has had on advancing drug discovery research. In vitro assays
typically use cells grown on two-dimensional hard plastic or glass
substrates which are not representative of the true in vivo cell environment. In
tissue cells interact with neighboring cells and with the extracellular matrix
or ECM to form a communication network that affects many cellular processes
including proliferation migration and apoptosis. In a simplified in vitro 2D
environment most of the tissue specific architecture and cell-cell communication
cues are lost therefore there is a need for advanced culture methods that better
mimic cellular function within living tissue. Three-dimensional cell culture
methods provide a matrix that encourages cells to organize into structures that
resemble the in vivo environment thereby developing normal cell to cell and cell
ECM interactions in an in vitro environment. What options do researchers have to aid in the formation of 3D cultured cells,
well one there are polymeric hard scaffolds typically composed of a
cross-linked plastic such as polystyrene. 2 there are biological scaffolds where
the cells are combined with and then suspended within a matrix of
extracellular matrix proteins typically found in vivo which provide structure
and support to the cells, and 3 the spheroid model which is also very
popular as it can mimic both tissues and tumors depending on the included cell
types pre-made spheroids or micro tissues can be provided to you in
microtiter plates to simplify or even eliminate the cell culturing process. If
costs are a hurdle for you or you want to have more control over your culturing
process there are two more options to create your own spheroid models.
First there is the hanging drop where cells are suspended in a 30 to 50 micro
liter drop which literally hangs below the bottom of the well. Cell aggregation
then occurs in the drop over time. Another popular method is to use micro
plates coated with an ultra low attachment or ULA surface. These plates
look and behave much like other round bottom plates. However when the cells are added the ULA coating prevents attachment to the bottom of the well
which promotes the cells to aggregate together into a spheroid structure, and
as the spheroids are already in the bottom of the micro plate well addition
of 3D overlays such as soft auger or Matrigel Matrix is easily accomplished. Once you have selected the culturing format you’ll need to think about how to
collect the information that will answer your questions.
Are you looking to monitor a cellular response to an experimental treatment or
are you focused on learning about detailed changes in the cellular
morphology. Or it could be a combination of both of these. Here are two methods
that can be used to collect information on your cells. They provide different
results so it really depends on the level of detail you are looking for to
achieve and answering your questions. Microplate readers which typically
employ a PMT based detector provide a rapid means to make Holwell measurements.
Experiments performed on 3D cultured cells and commonly run on micro plate
readers include cytotoxicity and signal transduction assays. On the other hand
microscopes and cell imagers increase the amount of information attained from
each experiment through the use of widefield microscopy by incorporating
magnification through the use of appropriate objectives. Unnecessary areas of the well are excluded from examination increasing the detail of the
image being viewed as well as the data generated. High content imagers provide
the greatest quantity and breath of imaging based data. Automation of the
imaging process in addition to the inclusion of high content analysis
software allows image acquisition and analysis to be performed in a high
throughput manner. If you need to combine multiple functionalities to obtain your
results there are options available to integrate microscopy with multimode
microplate reading can provide both phenotypic and quantitative results. From
a single precious sample multiple pieces of information can be captured but
quickly and easily. Depending on your research needs a
multifunctional instrument may be the right choice for you 3D cell culture as
an exciting and important experimental tool that has enabled researchers to
better simulate living tissue in an in vitro environment. You have many
options to choose from in regards to 3D culturing methods and the level of
detail required in the data collected while it can be difficult to navigate
through the available choices. Once you make your selection the results you can
get from using this technique will be truly invaluable.

One thought on “Biocompare Bench Tip – 3D Cell Culture

  1. Three-dimensional cell culture technology is a technology of cultivation and animal experiments between monolayer cells, which can simulate the in vivo environment, and show the advantages of the cell culture.

Leave a Reply

Your email address will not be published. Required fields are marked *