Download Applications of Human Amniotic Epithelial cells in Stem Cell Biology

Applications of Human Amniotic Epithelial cells in
Stem Cell Biology and Regenerative Medicine
Camilla Terracina, Isabel Hazan, Roni Greenberg
Rawan Damouni and Prof. Eliezer Shalev
Ha’Emek Hospital and Ruth and Bruce Rappaport Faculty of Medicine, Haifa, Israel
Laboratory for Research in Reproductive Sciences, Obstetrics and Gynecology, Ha’Emek Medical Center, Afula
Introduction
Regenerative medicine is a new field of medicine based on the plasticity of
stem cells. The placenta provides a non-controversial, abundant source of
cells, unlike the highly controversial embryonic stem cells. The placenta is
an organ tasked with providing nutrients to the fetus. It is composed of
three layers: the decidua, chorion and amnion. The decidua connects the
embryo and the mother, passing oxygen and nutrients through blood
vessels. The chorion is the outer of two membranes protecting the
developing human. The amnion, derived from the Epiblast, is the inner
membrane surrounding the fetus. Differentiation occurs eight days after
fertilization, with the formation of the Epiblast, and prior to gastrulation.
Research Procedure
Isolation
Culturing the cells in FCS medium
Characterize the cells
Protein expression profile by FACS
analysis and immunoflourecence
staining
Iug
Results
Protein Expression of Stem Cell Markers Oct-4 and SSE4
Figure 1: Structure of Placenta placenta consists two components: the fetal
surface contains; Chorion and amnion membrane, which develops from the same
blastocyst that forms the fetus, and the maternal surface (Decidua basalis),
which develops from the maternal uterine tissue. The placenta consists of the
mesodermal amniotic membrane (AM) covered by epithelial layer of cuboidal
and columnar cells (AE). The chorionic membrane (CM) consists of a
mesodermal layer and a layer of extravillous trophoblast (CT) cells. The amnion
membrane can be easily isolated.
Human amniotic epithelial cells (hAECs) are derived from the amniotic
layer of the placenta. There are three types of stem cells: totipotent,
pluripotent and multipotent, decreasing in differentiation capabilities
respectively
Figure 2: the application of hAECs.
pluripotent hAECs derived form the
epiblast (inner mass cells) 8 days after
the fertilization before gastrulation.
which offer the ability to differentiate
into the three germ layers: Ectoderm,
Mesoderm or Endodermic.
Figure 3:Embryonic stem
cells in the Inner Cell Mass
are pluripotent ,because
they have the ability to
differentiate into any of the
three germ layers that will
create the entirety of the
organism.
Objective of the study
To explore the differentiation potential of the hAECS
RESEARCH POSTER PRESENTATION DESIGN © 2011
www.PosterPresentatio
ns.com
% of Cells
A
100 90 80 70 60 50 40 30 20 10 0 4-Oct
SSE4
DO B
DAPI
OCT4
P3 Merged
P5 C
Marker
4-Oct
SSE-4
DAZL
Sp
Expected time
+
+
+
+
+
-
Figure 3: Protein Expression of OCT 4 and SSE-4 (Immunofluorescence staining, and FACS
analysis). (A) Percentage of cells expressing OCT 4 and SSE-4 at D0, P3 and P5 analyzed (B)
Expected and Actual results f germ markers in the cells. (C) Immunofluorescence staining of
passage 5 (P5) hAECs (488 - conjugated secondary antibody) .
Discussion
In the experiment that we conducted, we explored the pluripotency of hAECs to
differentiate into the three germ layers; the ectoderm, mesoderm and endoderm. It was
found that most of these pluripotent cells maintained their stem cell characteristics and
differentiation abilities while being cultured in Fetal Calf Serum, but the levels of SSE4
decreased slightly by passage 5, as a result of some of the cells displaying characteristics of
fibroblast cells by the end of the experiment, supporting our hypothesis that these cells do
have the differentiation capabilities of pluripotent cells while being cultured in the Fetal
Calf Serum.
Conclusion
These results demonstrate the promising potential in hAECs and their role in regenerative medicine,
as they are an abundant and ethical non-controversial source of undifferentiated stem cells.
References
1.Miki T, Lehmann T, Cai H, Stolz DB, Strom SC: Stem cell characteristics ofamniotic epithelial cells.
Stem Cells 2005, 23:1549–1555.
2. Ilancheran S, Michalska A, Peh G, Wallace EM, Pera M, Manuelpillai U: Stem cells derived from
human fetal membranes display multilineage differentiation potential. Biol Reprod 2007, 77:577–588.
3. EvronA, Goldman Sh and Shalev E: Human amniotic epithelial cells differentiate into cells
expressing germ cell specific markers when cultured in medium containing serum substitute
supplement. . Reproductive Biology and Endocrinology 2012, 10:108
Acknowledgements
We would like to thank our mentor, Rawan Damouni, PhD for kingly guiding us through our research project, and
thoughtfully answering all questions and concerns we had along the way . In addition, we would like to thank Prof.Eliezer
Shalev for hosting We would also like to sincerely thank the Wolf Family and Na’aleh for their generosity and donation.