Suppressing Teratoma Formation with PluiSIn 1

Marc Piercy, Parshwa Sabadra, Yuzana Mone, Matthew Low

Results

Background
Any treatment based on ES or iPS cells run the risk of tumor formation through
teratomas. Such tumors must be suppressed for stem cell based treatments to come to
fruition. A molecule, PluriSIn#1, was found to be toxic to human ES and iPS cells but
not differentiated cells. It is possible that PluriSIn#1 can suppress teratoma formation.
To test the effects of exposure to PlurSIn #1 on teratoma formation, ES and iPS cells
were spontaneously differentiated in culture then mixed 1:1 with undifferentiated
cells before being cultured in PlurSIn #1 for 48 hours. The expected result is to see no
teratoma formation among the mixture of cells that were treated with the PlurSIn #1.

Each mouse was

injected with same total
number of cells treated
with DMSO or
PluriSln#1 (side to side).

Purpose
To determine the effectiveness of PluriSIn#1 in
selectively killing undifferentiated cells.
To determine if Plurisin#1 can suppress teratoma
formation.

Methods
Add CSESSO2/2 Cells

Red fluorescence
means Oct4
positive cells

Add
treatments
DMSO

Stain for Oct4

Plurisin # 1

Wait 48 H

DMSO

Plurisin #1

Figure 1. Procedure to test if PluriSIn#1 kills undifferentiated Human stem cells. We expect to see that
exposure to PluriSIn#1 kills the majority of Oct4(marker for undifferentiated cells) positive cells, far
more than DMSO
Figure 2. Procedure to test if PluriSIn#1 can inhibit Teratoma formation. Expect that PluriSIn#1
treated cells will form fewer teratomas than DMSO treated cells

Figure 2

Culture cells and

allow to
differentiate
Human
iPS
Cells

Human
ES

Add
undifferentiated
cells in a 1:1
ratio. Add
PluriSIn 1 or
DMSO

10
Days

hiPSc

Plurisin 1

D
M
SO

Inspect for
teratomas

6 Weeks
Later

PluriSln#1 prevents
teratoma formation
from tumorigenic
undifferentiated cells.

Gut epithelium,
skeletal muscle,
and neural
rosette structures
derived from
ESC- and iPSCteratomas were
stained and
visualized.
Since these three structured indicate the presence of each germ
layer, we can conclude that the teratomas formed were
perfectly normal and pretty similar in shape and structure to
one another. Also, this confirms that no teratomas were formed
in mice injected with PluriSIn#1 treated cells because of the
effect of PluriSIn#1 on undifferentiated cells and not because of
any abnormalities in those specific undifferentiated cells.

Discussion

The table summarizes the results of the teratoma formation

experiments. We can see that all of the control cells, but none of
the PluriSIn#1 treated cells, form teratomas. We can thus
conclude that PluriSIn#1 selectively kills all undifferentiated
cells therefore blocking teratoma formation.

Acknowledgements

hESc

PluriSIn#1 kills nearly all undifferentiated human cells.

Wait 48 H
Inject the cells
into
immunodeficient
mice

CSES-SO2/3 cells after 48 hours of exposure to DMSO or

PluriSIn#1.
Fluorescence microscopy confirmed that Oct4 expressing
cells (red fluorescence) are eliminated after the treatment with
PluriSln#1.

After 6weeks, teratoma

formation is found only
in DMSO-treated cells,
but not in PluriSln#1
treated cells.

Section and
examine
teratomas

It could be possible to differentiate cells and then

introduce the cells into an organism with minimal risk
of teratoma formation using PluriSIn #1
Opens the possibility of treatments based on human
ES and iPS cells

We would like to express our sincere thanks and

deepest gratitude to Pavan Kadandale, Brian Sato, and
Jacob Milligan for all their mentorship, help and support
throughout this class and for the last 10 weeks.