ABSTRACT
Introduction:
The placental
extracellular matrix (ECM) dynamically remodels over
pregnancy and in
disease. How these changes impact placental
barrier function is poorly understood as there are limited
in vitro models of the
placenta with a modifiable stromal compartment to mechanistically investigate these extracellular factors. We developed a straightforward
method to incorporate
uniform hydrogels into standard
cell culture inserts for transplacental transport studies.
Methods:
Uniform polyacrylamide (PAA)
gels were polymerized within
cell culture inserts by (re)using the insert
packaging to create a closed, controllable environmental chamber. PAA pre-
polymer solution was added dropwise via a
syringe to the
cell culture insert and the
atmosphere was purged with an inert gas. Transport and
cell culture studies were conducted to validate the model.
Results:
We successfully incorporated and ECM functionalized
uniform PAA
gels to
cell culture inserts enable
cell adhesion and monolayer formation. Imaging and analyte transport studies validated gel formation and expected mass transport results and successful
cell studies confirmed
cell viability, monolayer formation, and that the model could be used transplacental transport studies. Detailed
methods and validation
protocols are included.
Discussion:
It is well appreciated that ECM biophysical and biochemical properties impact
cell phenotype and
cell signaling in many
tissues including the
placenta. The incorporation of a PAA gel within a
cell culture insert enables independent study of placental ECM biophysical and biochemical properties in the context of transplacental transport. These straightforward and low-
cost methods to build three dimensional cellular models are readily adoptable by the wider scientific
community.