Page 177 - ESHRE2019

Hydrogels and reproductive medicine
A new era in reproduction and regenerative medicine: Extracellular matrix hydrogels and coatings
•Further processing acellular scaffold:
• Sections or blocks for three‐dimensional cell culture systems.
• ECM hydrogels and coatings: in vivo and in vitro applications.
•ECM powders, converted in protein
monomeric components (solubilized):
Solubilize ECM
• At physiological temperature: spontaneous
Decellularized tissues and ECM hydrogels represent an interesting
reformation of the intramolecular bonds.
platform for in vitro and in vivo REPROTEN applications • Never done before for reproductive tissues.
(Christman et al,, 2012)
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27
Innovation
The Rabbit Model: Materials and Methods
Experimental Design
Whole organ decellularization.
Separation of endometrium via microdissection.
Nulliparous, non‐ proliferating (non‐ synchronous, NS) endometrium.
Reagent
Triton‐X100
Triton X‐100
DNase I solution
Concentration
1%
‐
Nulliparo
Duration
Day 1
PBS
SDS
H2Od
1X
0.10%
1 hr
18 hrs
‐
30 min
30 min
PBS
1X
5 hrs
Day 2
SDS
H2Od
0.10%
18 hrs
30 min
us, proliferating
Lyophilization and milling to NS and S ECM powder.
y
embryo, S) endometrium.
(s
nchronou
1
%
s
w
ith day 3
1
h
r
PBS
1X 1 hr
Only difference with previous protocol:
• Wash with 2μg/mL
DNase 1 solution.
(Campo et al., 2019) 28
2 μg/mL
1hr
PBS
1X
3‐4 hrs
NS S
Innovation n=2 n=2
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PRECONGRESS COURSE 11 I VIENNA, AUSTRIA – 23 JUNE 2019 175
Cycle 2 Cycle 1