Lions_Eye_Flipping_Book - page 7

Lions Eye Institute Newsletter |7
feed the cultures a media containing
D-Valine (an amino acid) rather than
the normal media with L-valine (same
amino acid but in a different chemical
configuration). Only epithelioid cell
(like RPE) are capable of using both the
L andD formof Valine while fibroblasts
can only use the L- isoformof the amino
Corneal endothelial cells (CEC)
are the single layer of cells on the back
surface of the cornea. These cells keep
the cornea clear by actively pumping
fluid out of the cornea as there is a
natural flow of fluid into the cornea.
Human CEC do not reproduce when
they are injured or damaged in the body
so if cells are damaged through disease
or surgical damage, the remaining cells
simply grow larger to fill in the space.
One of the challenges to grow these cells
in vitro is to convince them to multiply
and divide again. Fortunately, the ORC
collaborates with the SingaporeNational
Eye Center who has helped us with
specializedmedia that causes the cells to
grow and a secondmedia that causes the
cells to have the same shape and size that
they do on the intact cornea. In addition
to examining basic disease mechanisms
in these cells, they also could have a
future application. The long term goal
for these cells is to be able to grow them
for corneal transplants. Imagine that the
cells from one cornea could replicate
into enough cells for several transplants
rather than the current one donor for
one recipient transplant paradigm. It
will take many experiments, many
specialized growing conditions, and
many characterizations to satisfy FDA
regulations as well as extensive testing
but one day in the future this “science of
sight” hopefully will contribute to “the
gift of sight.
Adult human corneal endothelial
cells are beginning to attach and grow
on the culture plate. The large mass of
cells were removed from a donor cornea
by enzyme digestion, spun down using
a centrifuge and then placed into a cell
culture dish. The cells begin to settle on
the dish as well as grow away from the
mass of cells. The growing and moving
cells are characterized by the elongated
spindle shape.
Adult Human corneal endothelial
cells are forming a monolayer on the
culture plate. After the initial growth and
movement, the cells begin to enlarge. The
cells form sheets of connected cells with
a morphology similar to the intact cells
on the cornea.
This isantibodystainingof thenuclei of
adult corneal endothelial cells. The nuclei
of the cells are stained with florescent
antibodies to help characterize the cells
insuring that only corneal endothelial
cells are present.
Because of
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