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Growing
Cardiovascular
Tissues
In
Vitro

               Rich
Goodwin,
PhD

            Cell
Biology
and
Anatomy

         ...
A.
                        C.




              Lumen

        B.





 B.
                         D.


                 ...
Day
21
Cardiotube

Embryonic
Mouse
Cardiac
Myocytes
at
11
Days
of
Culture


TEMs
of
Tube
Cultured
Cardiac
Myocytes


Undergo
myofibrillogenesis
and
development
cell‐cell
juncMons





               ...
We
are
using
cardiotubes
to
test
mechanisms
of
cardiovascular
development

such
as
Tetralogy
of
Fallot
(Dr.
Kheradvar).

T...
*

     *




1
    2        3

Stage
25
AV
cushions
in
lumen
of
tube
scaffold





                   Day
2

The
All
New
and
Improved

  FLOWMASTER
3000!!!

   (John
Weidner,
USC)

Free
Edge

Tenascin
C
expression
and
localizaMon
regulated
by
Flow
and
RhoA

                           Hong
Tan

Flow
and
RhoA
can
regulate
the
material
properMes
of

developing
valves
as
measured
by
Atomic
Force
Microscopy



        ...
Valve
Conclusions

   •  Expression
of
Fibrous
ECM
proteins
can
be

      regulated
by
flow.

   •  We
hypothesize
that
the...
Coronary
Development


Many
Thanks:

                          JD
Pods
and
MJ
Yost
‐
USC
SOM

                                Bob
Price
‐
USC
IRF...
Key
Points:

• The
mechanisms
of
cardiovascular
development
can
invesMgated
using
3D

culturing
techniques.

• Development...
Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD
Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD
Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD
Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD
Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD
Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD
Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD
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Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD

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Growing Cardiovascular Tissues In Vitro, Rich Goodwin, PhD

  1. 1. Growing
Cardiovascular
Tissues
In
Vitro
 Rich
Goodwin,
PhD
 Cell
Biology
and
Anatomy
 USC
School
of
Medicine

  2. 2. A.
 C.
 Lumen
 B.
 B.
 D.
 D.
 C.

  3. 3. Day
21
Cardiotube

  4. 4. Embryonic
Mouse
Cardiac
Myocytes
at
11
Days
of
Culture


  5. 5. TEMs
of
Tube
Cultured
Cardiac
Myocytes

 Undergo
myofibrillogenesis
and
development
cell‐cell
juncMons
 Evans
et
al,
2003

  6. 6. We
are
using
cardiotubes
to
test
mechanisms
of
cardiovascular
development
 such
as
Tetralogy
of
Fallot
(Dr.
Kheradvar).
 Toxicity
Screening
(Dr.
Yost/Biotech)

  7. 7. *
 *
 1
 2 3

  8. 8. Stage
25
AV
cushions
in
lumen
of
tube
scaffold
 Day
2

  9. 9. The
All
New
and
Improved
 FLOWMASTER
3000!!!
 (John
Weidner,
USC)

  10. 10. Free
Edge

  11. 11. Tenascin
C
expression
and
localizaMon
regulated
by
Flow
and
RhoA
 Hong
Tan

  12. 12. Flow
and
RhoA
can
regulate
the
material
properMes
of
 developing
valves
as
measured
by
Atomic
Force
Microscopy

 14001
 No
Flow
 Flow
 Flow
w/
LPA
 Flow
w/
y‐compound
 60
 12001
 Young's
Modulus
(Pa)
 50
 10001
 40
 8001
 Force
(nN)
 30
 6001
 20
 4001
 10
 2001
 0
 1
 0
 500
 1000
 1500
 2000
 2500
 3000
 No
Flow
 Flow
 Flow
w/
y‐compound
 Flow
w/
LPA
 z
(nm)
 Hong
Tan,
Stef
Biechler,
 (P<0.05)
 and
Dr.
Dean
(Clemson)

  13. 13. Valve
Conclusions
 •  Expression
of
Fibrous
ECM
proteins
can
be
 regulated
by
flow.
 •  We
hypothesize
that
the
mechanical
forces
 created
by
pulsaVle
flow
deform
the
 cytoskeleton
and
acVvate
the
RhoA
pathway.
 •  Experimental
manipulaVon
of
the
RhoA
 pathway
does
regulate
Fibrous
ECM
expression
 and
localizaVon
in
developing
valves
 So,
if
you
want
to
make
stronger
valves
faster,
use
Flow
and
RhoA

  14. 14. Coronary
Development


  15. 15. Many
Thanks:
 JD
Pods
and
MJ
Yost
‐
USC
SOM
 Bob
Price
‐
USC
IRF

 My
Lab
 HJ
Evans‐Anderson
–
Alumnus
 Stefanie
Biechler
 Lorain
Junor
 MUSC
 Andrea
Roberts
 Roger
Markwald
 Samantha
SMnson
 Chip
Norris
 Hong
Tan
 USC
Engineering
 Northwestern:
 John
Weidner,
Arash
Kheradvar
 Bob
Dedman
 Clemson:
 NIH
Support
 Delphin
Dean
 Cobre
P20‐RR1634
(RRM)
 R21
HL080294
(RLG)
 RO1
HL086856
(RLG)
 NSF
RII
Bridge
 A.
Tsiaras
In
Life
Aug.
26th,
2005

  16. 16. Key
Points:
 • The
mechanisms
of
cardiovascular
development
can
invesMgated
using
3D
 culturing
techniques.
 • Developmental
mechanisms
are
likely
to
be
important
during
regeneraMon.
 Problems:
 • The
transiMon
from
capillary
to
larger
caliber
blood
vessels
has
been
 difficult
to
model.
 • Numerous
others.

 CollaboraMons:
 • Clinical
applicaMons.

 • Grateful
for
my
present
collaborators


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