eBook: Breakthroughs and Innovations in Immunology Research - 8

used to direct cell behavior, health, or growth dynamics.
However, it must also be noted that basement
membrane extracts are prone to lot-to-lot
diversity and can be hard to work with. Tailoring
the right concentration and protocol for any application
is critical to developing a successful model,
alongside product quality and consistency.
Cell culturing in two or three
dimensions
Developing the right hydrogel using basement
membrane extracts is challenging yet still crucial
for successful culturing. Depending on the concentration
or the integrity of the hydrogel formed,
cell behavior can differ significantly. For 2D cell culture,
whether it is for iPSCs or other primary cells,
a thin layer coating works best. Promoting surface
attachment and maintaining stem cell stemness is
critical, thus using a reduced growth factor extract
should be utilized to minimize differentiation influences
from intrinsic cytokines or growth factors.
Stem cell differentiation, however, may require the
addition of other stem cell growth factors or iPSC
growth factors. By mixing growth factors within
the gel before plating stem cells, differentiation
can be induced toward the targeted cell type.
3D cell culturing, on the other hand, takes advantage
of the gel-like properties of basement gel extracts
to provide the cellular structure needed to
form organ precursors, also known as organoids.
Instead of being derived from a cell line, organoids
are composed of various types of cells that match
its starting material composition, providing an in
vitro 'snapshot' of cell diversity and structure. This is
extremely useful in immunology disease modeling,
especially for research looking deeper into cell-tissue
or more macro-level interactions. In most cases,
when culturing organoids, tissue fragments are
mostly used as a starting material. Depending on
Figure 1. Differentiation and expansion of (A) induced
pluripotent stem cells (iPSCs) into (B) myocardial cell
precursors after induction.
the desired organoid type, both thin layer and thick
layer gels are sufficient to provide the optimal environment
for 3-dimensional cellular growth. This
includes mouse intestinal organoids, hepatobiliary
organoids, and lung tracheal organoids.
Certain organoids may be harder to culture, such
as human tumor organoids. By mixing the starting
material with the basement membrane extract before
plating and gelling, improved organoid structure
can be observed. A denser extracellular matrix
layer and thicker gel layer provide better scaffolding
for cell attachment and growth. Similarly,
organoid growth factors can be added to further
help promote growth and accurate cell diversity.
Disease modeling-Angiogenesis
and xenografting
Whereas organoids provide a good cellular-level
precursor model for immunology research, baseFigure
2. Culturing of (A) mouse intestinal organoids, (B) mouse
liver ductal organoids, and (C) mouse airway organoids in a 70%
Matrigengel Matrix GFR gel.
8

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eBook: Breakthroughs and Innovations in Immunology Research

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