(188ah) Engineering Pancreatic Islet Organoids from Human Pluripotent Stem Cells

Poster
title

Engineering
Pancreatic Islet Organoids from human Pluripotent Stem Cells

Author
list

Humphrey,
Nadine; Wiegand, Connor; Carnaval, Izzy; Richardson, Thomas; Banerjee, Ipsita

Brief
description of research (no longer that 500 words)

This project explored the role of human pluripotent stem
cells as a potential cell source for generating pancreatic islet organoids for regenerative
cell therapy of Type 1 Diabetes.

Type 1
Diabetes (T1D) is a chronic disease that occurs when the body cannot produce
enough insulin. Over time, T1D can cause damage to the heart, blood vessels,
eyes, kidneys, and nerves. Current long-term treatments require cumbersome
daily glucose monitoring and insulin injection.  Transplantation of isolated
pancreatic islets into a diabetic patient offers a more permanent treatment but
has limited applicability because of scarcity of donor islets. Derivation of
pancreatic islets from human pluripotent stem cells (hPSCs) will have the
potential to overcome this scarcity and hence will have a significant effect in
cell therapy of diabetes.  

In this
project, we used alternative strategies to generate pancreatic islet ‘organoids’
from human pluripotent stem cells, targeting to reproduce the three-dimensional
(3D) spheroid structure of the derived islets. We further compared the
islet-specific function of these derived islet organoids, to evaluate the best
strategy for organoid engineering. The Banerjee lab has an established protocol
for derivation of pancreatic beta-like cells from hPSCs under adherent (2D)
culture. Using this protocol, we differentiated the hPSCs to a pancreatic
progenitor stage (hPSC-PP), where the hPSCs start expressing PDX1, a robust marker
for pancreatic lineage. At this point, the hPSC-PP cells were harvested and
aggregated into 3D spheroids using 3 alternate platforms: amikagel, Agarose and
ultra-low adhesive plates.

Notably,
these 3 platforms have progressively reduced cell binding properties: amikagel
has weak binding, agarose typically has no binding and ULA is specifically
treated to prevent cell binding. The hPSCs successfully aggregated into stable
spheroids on each of these platforms, with comparable initial aggregation
dynamics. However, their long-term condensation dynamics was significantly
different, which eventually affected the cell fate, and hence the
islet-organoid function. For example, on Amikagel the organoids condensed
significantly, whereas in ULA there was obvious cell growth. From the
immunostaining, organoids on ULA observed the largest secretion of PDX1 and INS,
another robust pancreatic lineage marker, but all of the platforms had little
NKX61 secretion. Interestingly, all the organoids were functional, and secreted
insulin in response to blood glucose levels, with the organoids derived on
Amikagel exhibiting highest glucose responsive insulin release. In the next
step we are implanting these organoids under the kidney capsule of mouse models
to evaluate their in-vivo potential.   

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