Direct labeling.

The most common contrast agents used for direct label-ing

3335
are nanoparticles or radionuclides
. Nanoparticles suitable for MRI
include superparamagnetic iron oxide (SPIO) nanoparticles (i.e.,
ferumoxides, ferucarbotran-1, ferumoxytol), gadolinium-filled micro36
capsules and liposomes, perfluorocarbon nanoparticles and manga-nese37
based particles (Table 1). Advantages of nanoparticle-based MRI cell
tracking are a strong signal, allowing high-resolution visualization of the

migration and homing of injected cells, as well as relative ease of cell

labeling. SPIO nanoparticles can also be detected by ultrasound, although
38
this approach is not widely used . SPIO-based labeling has been used
clinically to track transplanted cells in the brain for up to 7 weeks by
39,40
MRI
. Labeling cells with perfluorocarbon nanoparticles is a promising
approach that is being tested in clinical trials.

wing
detec
D tion
ir
of as
ec
few
t
as
4
la
~10
be
6
li
10
n
cells/
g
voxel
4
w
.
it
Radi
h
onucl
ra
ide
di
labeli
o
ng is
n
readi
uc
ly
li
quant
de
ified
s
and
fo
can
r
offer
S
even
P
great
E
er
C
sensi
T
tivity
an
than
d
MRI,
P
but
E
halfT
lives
i
are
m
relati
ag
vely
in
short
g
, and
ha
the
s
limit
ex
ed
ce
spati
ll
al
en
resol
t
ution
se
of
ns
clinic
iti
al
vi
PET
ty
and
co
SPE
m
CT
pa
requi
re
res
d
imag
w
e coit
locali
h
zatio
M
n
R
with
I,
CT
C
or
T
MRI.
an
More
d
over,
ul
conc
tr
erns
as
regar
o
ding
u
radia
n
tion
d
expo
be
sure
ca
may
us
limit
e
these
th
appr
e
oach
ba
41
ck
es .
gr
Direc
o
t
u
labeli
n
ng
d
with
si
radio
g
isoto
na
pes
l
such
is
as
111
lo
In
w,
dium
al
(In)lo

o
x

ce

A lls
l .

So
lu
bl
e
ra
di
oi
bele
so
d
to
anti
pe
bodi
es s
m
ay
be
le
ss
pr
on
19
e
to
thi
s
art
ifa
ct;
to
ou
r
kn
o
wl
ed
ge
,
HSV1-tk
th
eir
Reporter
up
proteins
ta
ke
by
m
ac
ro
ph
Ferritin
ag
es
ha
LRPs
no
t
be
en
st
ud
ie
d.
Se
co
nd
,
co
nt
ra
st
ag
en
ts
ar
e
dil
ut
ed
as
ce
lls

divid
e,
result
ing in
a
gradu
al
disap
pearance
of the
signa
49

51

.
Third
,
imagi
ng
signa
ls can
be
diffic
ult to
distin
guish
from
back
groun
d.
For
exam
ple,
SPIO
nano
partic
les
are
detec
table
in
MRI
as
areas
of
decre
ased
signa
l
inten
sity,
and
simil
ar
signa
l
voids
can
be
cause
d by
iron
depos
itions
in the
form
of
hemo
sideri
n
from
old
hemorrha
52

ge .
Finall
y, all
contr

a
s

I
n

la
be
li
n
g.
In
in
di
re
ct
la
be
li
ng
,
ce
lls
ar
e
ge
ne
tic
all
y
m
od
ifi
ed
to
ex
pr
es
s
re
po
rt
er
ge
ne
s
en
co
di
ng
pr
ot
ei
ns
th
at
ge
ne
ra
te
i
m
ag
in
g
si
gna
ls,
of
te
n
up
on
in

terac
tion
with
a
mole
cular
prob
e or
subst
rate
that
is
taken
up
by
cells.
Repo
rter
gene
s
incor
porat
ed
into
the
geno
me
are
prop
agate
d by
daug
hter
cells,
and
the
imag
ing
signa
l is
stoic
hiom
etrically
relat
ed to
live
cell
mass
.
Epis
omal
expr
essio
n of
repor
ter
gene
s is
also
possi
ble,
but
diluti
on of
episo
mal
vecto
rs

makes them
unsuitable for longterm track-

ing of mitotically active cells. Reporter-gene

imaging is widely used in animal research, but
clinical applications for use in tracking cells to
treat cardiovascular or central nervous system
diseases have been limited by concerns about

18

F-FDG
99m

TcHMPAO

111

In oxine

the safety of genomic integration and about

potential immune responses to some of the
foreign reporter proteins.
A variety of reporter proteins detectable by
different imaging modalities have been developed. Firefly luciferase, detectable by bioluminescence imaging, is often used in small3
animal research ; however, absorption of the
emitted light precludes its use in most largeanimal and human studies. For large-animal
and clinical applications, there are several
potential options.
The iron-storage protein ferritin is detectable by MRI as voxels with reduced signal

SPIO

5658

intensity

. Because ferritin is ubiquitous in

18

F-FHBG

Reporter

Katie Vicari/Nature

gene

Group
gPublishin

Vector

Brain

Figure 2 Tracking cell fate by noninvasive imaging

requires either direct or indirect labeling.
(a) Direct labeling. Exogenous labeling with either
MRI-based contrast agents or radioprobes for
19
PET or SPECT imaging. Cells take up SPIO or F
nanoparticles (Fnp) primarily through endocytosis,
99m
111
whereas
TcHMPAO or
Indium oxine are
lipophilic and pass through cell membranes by
passive diffusion. FDG is a glucose analog and is
taken up though glucose transporter channels on
cells. Small molecules can attach to cell surface
markers or enter cells through channels.
(b) Indirect labeling. Reporter genes introduced
into the genome express surface proteins,
channels, storage proteins or enzymes that
are detectable or that bind detectable probes.
HSV1-tk, herpes simplex virus thymidine kinase;
NIS, sodium iodide symporter; LRP, lysine-rich
protein; SPIO, superparamagnetic iron oxide
nanoparticles.

Parkinson's
disease

Intracerebral
injection

axial MRI,
PET overlay
18

Fetal
dopaminergic
neurons

FDOPA

Brain

Trauma

Cortex and
white matter
near lateral
ventricle (V)

Axial MRI
SPIO

Intracerebral
injection

Neural stem
cells

I
m
m
a

0% ID/g

H
L

L
Baseline

Day 1

Day 7
G

Pre

3 years PI

R L
Pre

Day 2

D1

D7

*
Figure 3
Examples of
clinical imaging
used to identify
and track
labeled cells in
the body. (a)
Coronal SPECT
images of chest
and upper
abdomen
obtained 18 h
after
intracoronary
(IC) versus
transendocardial
(TE) delivery of
about 10
99m

TcHMPAO
+

CD34
hematopoietic
stem cells in
patients with
nonischemic
dilated
cardiomyopathy.
IC route of
administration
clearly shows
less retention of
cells in the
myocardium
compared to TE
route with both
delivery
techniques
showing the
distribution of
+

CD34 cells in
liver and spleen
(IC > TE). This
comparison
demonstrates
the value of
short-term
labeling of a cell
product and the
value in
assessing the
cell delivery
route. Figure
from ref. 73
reprinted with
permission. (b)
PET/CT fused
axial multimodal

D14

D21

imaging of pig
chest performed
on clinical
scanner. MSC
transduced by
adenovirus
containing
cytomegalovirus
promoter driving
HSV1-tk
reporter gene
implanted with
matrigel into
porcine left
ventricle (LV)
myocardium
after
thoracotomy
(long arrows). T
is the chest tube
inserted during
8

surgery. 10
human MSCs
injected into the
myocardium
were visualized
(short arrows) 4
h after
intravenous
infusion of 9-(418F-3[hydroxymethyl]
butyl)-guanine,
a thymidine
analog that is
phosphorylated
by HSV1-tk
reporter
following uptake
by cells. % ID/g
is percentage
uptake per gram
of tissue. Figure
from ref. 32
reprinted with
permission. (c)
Coronal MRI
performed at 3 T
of liver from a
diabetic patient
baseline, day 1
and day 7
following portal
vein infusion of
5

4.81 10
SPIO-labeled
islet equivalents
demonstrating
hypointense
voxels
throughout the
liver (white
arrows) that
cleared rapidly
over 24 weeks
of imaging
follow-up. The
number of
hypointense
areas in liver
decreased by
about 5060%
in the first week
after infusion,
representing
rapid clearance
of the
transplanted
islets despite
immunosuppres
sion of the
patient. Figure
adapted from
ref. 104 with
permission. (d)
Coronal and
axial, T2weighted, 1.0 T
MRI following
implantation of
fetal neurons in
a patient with

Parkinsons
disease.
Coronal MRI
displays the cell
transplantation
needle track
(arrowheads)
following
injection of ~3.2
6

18

10 cells. F
fluorodopa PET
parametric map
fused with MRI
performed
before (PRE)
and 3 years post
implantation (PI)
of fetal cell
neurons that
matured with
time that
resulted
in
increased
uptake of
the
dopamine
analog by
the
innervated
cells in the
putamen.
Figure
adapted
from ref.
76 with
permission
. (e) T2weighted,
axial, 3 T
MRI before
(PRE) and
day 1 after
implantatio
n in the left
temporal
lobe of
SPIOlabeled
autologous
neural
stem cells
in patient
with
traumatic
brain

injury (* is
site of injury).
Magnified
serial T2*weighted,
axial MRI
performed on
days (D)
1,7,14,21
post
implantation
(PI) of
labeled cells
are shown.
Four
hypointense
areas (black
arrows) were
observed on
PI days
1,7,14, 21
around lesion
site (*)
followed by
migration of
cells (white

arrowhead
and arrows)
along the
border of the
damaged
area.
Hypointense
areas where
labeled cells
were injected
cleared over
time (D14,
D21), and by
week 7 PI,
areas were
no longer
visible on
MRI. Figure
adapted from
ref. 40 with
permission.

m
o
st
or
g
a
ni
s
m
s,
it
m
a
y
b
e
p
o
ss
ib
le
to
u
se
it
cl
in
ic
al
ly
.
H
o
w
e
v
er
,
M
R
I
se
nsi
ti
vi
ty
to
fe
rr
iti
n
is
lo
w
er
th
a
n
to
S
P
I

O
nano
parti
cles
(wor
k of
A.V.
N.,
C.E.
M.
and
colle
ague
46
s) .
He
rpes
simpl
ex
virus
thym
idine
kinas
e
type
1
(HS
V1tk) is
frequ
ently
used
for
PET
imag
ing
in
large
anim
32,
als
59
,
and
it has
been
incor
porated
into
sensi
tized,
cytol
ytic
Tcells
to
imag
e
meta
stase
s in
patie
nts
with
gliob
lasto
ma
using
the
ganci
clovi

ap
A pl
n ic
T ati
h on
of
T
di
h
re
C ct
h la
be
ls
is
re
al
ti
m
e
M
RI
gu
id
an
ce
of
ce
ll
de
li
ve
ry
,
as
no
ot
he
r
i
m
ag
in
g
m
od
ali
ty
pr
ov
id
es
as
pr
ec
is
e
an
at
o
m
ic
al
in
fo
r
m
ati
on
ab
ou

t cell
migr
ation
and
short
-term
biodi
strib
ution
in
vivo
9

. In
our
view,
indir
ect
label
ing
appr
oach
es
are
best
used
when
it is
nece
ssary
to
moni
tor
the
fate
of
impl
anted
cells
over
week
s,
mont
hs or
years
in
order
to
ensur
e that
engr
aftm
ent
has
occu
rred
in
the
targe
ted
tissu
e. An
addit
ional
adva
ntage
of
indir
ect
label
ing is
the

lls
I ca
m n
al
I so
n be
I as
m se
ss
C ed
eP us
r in
g
re
p
or
te
r
ge
ne
s
be
ca
us
e
th
e
i
m
ag
in
g
si
g
na
l
is
pa
ss
ed
to
da
u
g
ht
er
ce
lls
8,

74

.
T
he
m
os
t
co
m
m
o
n
ap
pr
oa
ch
,
de
m
o
ns
tr
at
ed
in
an
i
m

als,
is to
imag
e
seria
lly
and
to
esti
mate
grow
th
kinet
ics
from
the
rate
of
chan
ge in
popu
latio
n
size.
Repe
titive
,
long
term
(up
to 5
mont
hs)
PET/
CT
studi
es of
MS
Cs
expr
essin
g
HSV
1-tk
were
cond
ucte
d
with
a
porci
ne
mod
el of
myo
cardi
al
infar
ction
.
Cell
proli
ferat
ion
peak
ed at
33
35
days
after
injec
tion
in
peri
-infa
rct
regio

ag
I in
m g
di
ff
er
en
tia
ti
on
of
tr
an
sp
la
nt
ed
ce
lls
ha
s
re
ce
iv
ed
re
lat
iv
el
y
lit
tle
att
en
ti
on
,
an
d
th
er
e
ar
e
in
te
re
sti
ng
op
po
rt
un
iti
es
in
th
is
ar
ea
.
Pr
o
m
isi
ng
ap
pr
oa
ch
es
in
cl
ud

e
genet
icall
y
integ
rated
stage
speci
fic
prom
oters
and
label
ed
small
mole
cules
that
inter
act
with
stage
speci
fic
cellu
lar
targe
ts.
The
latter
appr
oach
is
well
suite
d to
PET
and
SPE
CT if
the
small
mole
cule
can
be
sligh
tly
modi
fied
by
addit
ion
of a
radio
label.
For
exam
-ple,
the
PET
agent
18

FDOP
A
was
used
to
moni
tor
matu
ratio
n of

I
m
ag
in
g
th
e
h
os
t
en
vi
ro
n
m
en
t.
D
es
pi
te
its
m
an
y
li
m
ita
ti
on
s
fo
r
tr
ac
kin
g
tr
an
sp
la
nt
ed
ce
lls
,
cli
ni
ca
l
i
m
ag
in
g
is
in
di
sp
en
sa
bl
e
in
re
ge

nerat
ive
medi
cine
for
unde
rstan
ding
the
host
envir
onm
ent.
Imag
ing
can
answ
er
man
y
speci
fic
quest
ions
abou
t
tissu
es
both
for
pretr
eatm
ent
plan
ning
and
for
longi
tudin
al
evalu
ation
of
respo
nse
to
thera
py
(Tab
le 2).
For
exam
ple,
what
is the
statu
s of
the
targe
ted
regio
n
with
regar
d to
various
path

I pr
m ov
H e
e tr

I ue
m
m
yo
ca
rd
ial
re
ge
ne
rat
io
n:
(i)
an
in
cr
ea
se
in
th
e
vo
lu
m
e
of
vi
ab
le
m
yo
ca
rd
iu
m
wi
thi
n
th
e
in
fa
rct
zo
ne
;
(ii
)
str
uc
tu
ral
int
eg
rat
io
n
of
th
e
ne
w
m
yo
ca
rd

ium
with
the
host
tissue
,
inclu
ding
restor
ation
of
myoc
ardial
fiber
archit
ectur
e;
and
(iii)
functi
onal
integr
ation
of the
new
myoc
ardiu
m
with
host
myoc
ardiu
m
that
is,
synch
ronou
s
contr
actio
n of
new
and
old
host
myoc
ardiu
m
witho
ut
cond
uctio
n
delay
or
arrhy
thmia
s.
Medi
cal
imagi
ng is
essen
tial in
the
evalu
ation
of
cardi
ac
repair

ud

C ie
l s
ha
I
ve
m
al
so
re
ve
al
ed
th
at
re
te
nt
io
n
in
th
e
is
ch
e
m
ic
m
yo
ca
rd
iu
m
de
pe
nd
s
on
th
e
de
li
ve
ry
m
et
ho
d.
In
th
e
he
m
at
op
oi
eti
c
st
e
m
ce
ll
tri
al
di
sc
us
se
d
ab
ov
e

, at
18 h
after
the
proc
edur
e,
myo
cardial
reten
tion
was
high
er in
the
trans
endo
cardi
al
grou
p
than
in
the
intra
coro
nary
grou
p.
Tran
send
ocar
dial
deliv
ery
also
corre
lated
with
great
er
impr
ove
ment
in
ventr
icula
r
funct
ion 6
mont
hs
after
infus
ion.
In a
5year
follo
w-up
study
80

,
cell
thera
py
was
assoc
iated
with
incre
ased

Table 2
Interrogating
the host
microenviron
ment by
clinical
imaging
Tissue characteristics
Anatomy (structure, volume loss, vasculature)
Vascular permeability
Pathology (e.g., mass)
Compositions (e.g., fibrosis)
Inflammation (acute, subacute, chronic)
Physiology and function

MRI
++++
+++
++++
+++
+++
++++

MRSI

++

Imaging techniques
X-ray angiography CT
SPECT/CT
++++
+++
+

++
+

++
+

++
++

++

++
+++

PET/CT
+
+
++
++
++
+++

Ultrasound
+++
++
+
++

++

Perfusion
Metabolism
Oxygenation
Cell viability
Cell identity

++++

+++

+++

++
++++
++
++++
++++

MRSI, magnetic
resonance
spectroscopy
imaging. MRI
techniques include
blood oxygenation
level dependent
contrast, diffusion
tensor imaging,
MR elastography,
dynamic contrast
enhance MRI.
not useful; +
limited to ++++
most useful.

d
es
ig
ni
n
g
s
y
st
e
m
s
th
at
in
te
gr
at
e
bl
o
o
d
fl
o
w
d
y
n
a
m
ic
s
w
it
h
ti

ssue
mec
hani
cs.
S
yste
mati
c
com
paris
ons
of
diffe
rent
rout
es of
adm
inist
ratio
n
have
been
carri
ed
out
in
ani
mal
mod
els.
MRI
and
PET
ima
ging
of
acut
ely
infar
cted
rats
sho
wed
that
dire

+++

c
t

li
v
er
a
n
d
s
pl
e
e
n

sults
and
sugg
ests
strat
egie
s for
impr
ovin
g
ther
8
apeu
tic
2
. effic
T acy.
Rete
h
ntio
e
n
u
se has
of been
incr
i
m ease
d by
a
gi injec
ting
n
cells
g
to in a
re hydr
ogel
v
mad
e
al e of
th natu
ral
e
lo or
w synre theti
te c
nt mate
io rials.
This
n
of appr
oach
tr
has
a
been
n
teste
s
pl d in
pigs
a
nt usin
g
e
hum
d
an
c
el MS
Cs
ls
pr expr
essi
o
vi ng
HS
d
es V1tk
a
and
p
PET
o
ss /CT
ib ima
le ging
32
e
x
(Fig.
pl 3).
a
Surp
n
risat ingl
io y,
n
myo
fo card
r
ial
p
radi
o
otra
or cer
cl
18
in ( F
ic HB
al G)
re upta

k
e

an
i
m
al
s
Q an
d
u hu
m
an
O
nA
s

7,
87
,8
8

.
M
yo
ca
rd
ial
fi
be
rs
ha
ve
a
co
m
pl
ic
at
ed
he
lic
al
str
uc
tu
re
th
at
is
cr
iti
ca
l
fo
r
ef
fi
ci
en
t
co
nt
ra
cti
le
an
d
co
nd
uc
ti
ve
fu
nc
ti
on
of
th
e
he
ar
t.
D
TI
tr

actog
raph
y has
show
n a
smoo
th
trans
ition
in
fiber
orien
tatio
n
from
epica
rdiu
m to
endo
cardi
um
in
the
healt
hy
heart
and
sever
e
disru
ption
of
myof
iber
archi
tectu
re
after
infar
ction
in
differ
-ent
speci
es
(hum
an,
shee
p,
rat)

.
Preli
mina
ry
DTI
resul
ts in
a
muri
ne
infar
ct
mod
77

el
sugg
est
that
this
techn
ique
has
pote
ntial
for
asses
sing
struc
tural
integ
ratio
n and
align

m d
e isl
et
F s,
i no
I co
rr
n el
I ati
m on
w
I
as
n
se
en
be
tw
ee
n
th
e
nu
m
be
r
of
in
je
ct
ed
isl
et
s
an
d
th
e
S
PI
O
si
gn
al
on
M
RI
i
m
ag
es
10
3

.
H
o
w
ev
er,
a
su
bs
eq
ue
nt
tri
al
us
in
g
fe
ru
ca
rb
ot
ra
nla
be
le
d
isl
et
s

dem
onstr
ated
a
60%
decre
ase
in
graft
volu
me
one
week
after
trans
plant
ation
in
eight
patie
nts
by
MRI
104

(Fig.
3).
Alth
ough
no
independ
ent
meth
od
was
used
to
confi
rm
this
findi
ng,
signi
fican
t Cpepti
de
level
s and
nearnorm
al
HbA
1c
value
s
were
achie
ved
in
these
patie
nts
with
a
subst
antia
l
redu
ction
of
insul
in
dose,
sugg
estin
g
that
the
rema
in-

B
o
x
3
A
f
r
a
m
e
w
o
r
k

npg

2014 Nature America, Inc. All rights reserved.

f
o
r

s
p
e
c
i
f
i
c
s
t
e
p
s
f
o
r

i
m
p
l
e
m
e
n
t
i
n
g

r
e
s
e
a
r
c
h
e
r
s

i
m
a
g
i
n
g

w
i
s
h
i
n
g

i
n
r
e
g
e
n
e
r
a
t
i
v
e
m
e
d
i
c
i
n
e
S
o
m

t
o
i
n
c
o
r
p
o
r
a
t
e
i
m
a
g
i
n
g
i
n

t
o

ce

T ll
e ty

pe

R
s
e (b
S
t

et
a
D ce
lls
o
,
I al
n ph
i a
n ce
lls
S ,
c de
b lta
e ce
lls
an
H d
o pa
T nc
h re
ati
T c
h po
ly
pe
pti
de
pr
od
uc
in
g
ce
lls
)

10

.
La
be
lin
g
of
thi
s
rat
he
r
lar
ge
cl
us
ter
of
ce
lls
is
no
nsp
ec
ifi
c
an
d
oc
cu
rs

only
by
diffus
ion.
In the
case
of
SPIO
nano
partic
les,
12
48 h
are
requi
red to
achie
ve a
conce
ntrati
on of
212
pg/ce
103,

ll

108,11
1

(wor
k of
A.M.
and
colle
agues
). In
vivo,
quant
ificati
on of
SPIO
label
ed
grafts
is
comp
licate
d by
the
prese
nce
of
susce
ptibil
ity
artifa
cts,
the
inabil
-ity
to
distin
guish
singl
e
islets
from
islet
cluste
rs
and
low
signa
l-tonoise
ratios
betw

e
eS

pr
o
vi
de
p
os
iti
ve
co
nt
ra
st.
A
st
u
d
y
in
ra
ts
sh
o
w
ed
hi
g
h
p
os
iti
ve
co
nt
ra
st
fr
o
m
S
PI
O
la
be
le
d
isl
et
s
o
n
th
re
edi
m
en
si
o
na
l
d
ua
l
ec
h
o
ul
tr
as
h

ort
echo
time
imag
es
and
supp
ressi
on of
the
sign
al
from
liver
and
smal
l
vess
els,
allo
wing
for
auto
mati
c
quan
tifica
tion.
Qua
ntifi
catio
n of
hype
rinte
nse
pixel
s
corre
lated
with
the
num
ber
of
injec
ted
IEQs
due
to
the
unif
orm
supp
resse
d
back
grou
nd
and
high
contr
ast
and
appe
ars
to be
supe
rior
to
stan
dard

i im
m ag
G in
a g
m

L od
o

ali

T tie
h s
F
i

co
ul
d
pr
ov
id
e
co
ns
id
er
ab
ly
m
or
e
in
fo
r
m
ati
on
re
ga
rd
in
g
gr
aft
fat
e
th
an
a
si
ng
le
ap
pr
oa
ch
.
To
th
at
en
d,
a
tri
m
od
al
mi
cr
oc
ap
su
le
fo
r
isl
et
la

belin
g
containin
g
gold
nano
partic
les
functi
onali
zed
with
DTD
TPA
(dithi
olate
d
dieth
ylene
triam
inepe
ntaac
etic
acid)
and
gadol
inium
chela
tes
was
devel
oped
for
coencap
sulati
on
with
islets
using
prota
mine
sulfat
e as a
clinic
algrade
algin
ate
cross
linker
.
Micr
oenca
psula
tion
decre
ased
islet
reject
ion
due
to the
prese
nce
of a
semiperm
eable
mem

I
n

us
io
n
of
pe
rfl
uo
ro
ca
rb
on
e
m
ul
si
on
s
in
al
gi
na
te
co
ns
tr
uc
ts
fo
r
isl
et
en
ca
ps
ul
ati
on
11
8,
11
9

al
so
all
o
w
s
fo
r
th
ei
r
de
te
cti
on
by
19

F
M
RI
,
ul
tr
aso
un
d
an
d
C

T (in
the
case
of
perfl
uoro
octyl
brom
ide)
and
does
not
alter
the
perm
eabil
ity of
the
caps
ules
or
affec
t islet
funct
ion.
Thes
e
new
appr
oach
es
not
only
impr
oved
islet
prote
ction
but
also
allo
wed
for
MRI
of
trans
plant
ed
islets
with
posit
ive
contr
ast
and
for
islet
infus
ions

u
nI
n

d and
engra
fted
islets
are
still
unde
r
devel
opme
d nt, as
ist are
ri
prob
bu es
ti
that
on are
,
quant
su itativ
rv
e,
iv
nont
al
oxic
an and
d
long
fu
retai
nc ned
ti
by
on islet
in
cells.
th
Furth
e
ermo
sh re,
or the
t
descr
an ibed
d
meth
lo
ods
ng label
te
islet
r
cells
m. and
H
are
o
not
w
speci
ev fic
er, for
al
beta
go cells.
rit
Ima
h
ging
m
brai
s
n
fo
repa
ir.
r
pr Acut
e
ec
brai
is
n
e
injur
qu y
an from
strok
tif e
ic
and
ati chro
on nic
neur
of
odeg
in
ener
fu
ative
se dise

a
s

In
T pa
h tie
nt
s
wi
th
m
ul
tis
ys
te
m
at
ro
ph
y,
in
fu
si
on
of
un
la
be
le
d
au
to
lo
go
us
M
S
C
s
in
to
th
e
ca
ro
ti
d
ar
te
ry
le
d
to
in
cr
ea
se
d
up
ta
ke
of
18

FF
D
G
in
va
ri
ou
s
ar
ea
s
of
th
e
br

ain
comp
ared
to
the
contr
ol
grou
p and
impr
ovement
in
funct
ional
neur
ologi
cal
outco
me
over
a
perio
d of
1
1

year
33

.
Of
conc
ern,
howe
ver,
diffu
sionweig
hted
MRI
detec
ted
areas
of
micr
oinfarcti
on,
presu
mabl
y due
to
MSC
clum
ping
in
vasc
ulatu
re.
Long
itudi
nal
imag
ing
studi
es
are
an
essen
tial
part
of
cell
thera
py
clinic
al
trials
to
moni

t
o

ce
ll
E pr
f oli
fe
I
rat
m
io
n
an
d
di
ffe
re
nti
ati
on
ha
s
no
t
be
en
de
m
on
str
at
ed
in
th
e
hu
m
an
br
ai
n.
A
m
aj
or
ob
st
ac
le
ha
s
be
en
th
e
de
ve
lo
pm
en
t
of
su
ita
bl
e
pr
ob
es
th
at
ca
n
cr

oss
the
blood
brain
barrie
r.
Poten
tial
indic
ators
of
integr
ation,
such
as
brain
plasti
city
and
cell
differ
entiat
ion,
can
be
asses
sed
by
functi
onal
MRI

37

or
MR
spect
rosco
138

py

,139

,
respe
ctively
. Remyeli
natio
n
after
neura
l
stem
cell
impla
ntatio
n in
patie
nts

Box 4
Recom
menda
tions
for
preclini
cal
evaluat
ion of
imagin
g
metho
ds
I
f
i
m
a
g
i
n
g
i
s
t
o
b
e
u
s
e
d
i
n
a
c
e
l
l
t
h
e
r
a
p
y
c
l
i
n
i
c
a
l

tria
l, it
is
ne
ces
sar
y
to
co
mp
ile
ap
pro
pri
ate
dat
a
for
pre
se
nta
tio
n
dur
ing
pre
IN
D
dis
cus
sio
ns
wit
h
the
reg
ula
tor
y
ag
en
cy.
Th
es
e
dat
a
sh
oul
d
incl
ud

e
I
n

n
d
i
n
c
l
u
d
e
d
e
t
e
r
m
i
n
a
t
i
o
n
o
f
l
a
b
e
l
i
n
g
e
f
f
i
c
i
e
n
c
y
;
l
a
b
e
l
c
o
n
c
e
n
t
r
a
t
i
o

n;
ra
te
of
c
ell
d
e
at
h;
s
h
or
t
a
n
d
lo
n
g
er
te
r
m
pr
oli
fe
ra
ti
o
n
c
a
p
a
cit
y;
di
ff
er
e
nt
ia
ti
o
n
c
a
p
a
cit
y;
m
ig
ra
ti
o
n
c
a
p
a
cit
y;
i
m
m
u

n
o
I

A
n

tF

e
s
t
i
m
a
t
e
o
f
t
h
e
d
i
l
u
t
i
o
n
r
a
t
e
o
f
t
h
e
l
a
b
e
l
o
v
e
r
m
u
l
t
i
p
l
e
c
e
l
l

d
i
v
i
s
i
o
n
s
c
a
n
b
e
p
r
o
v
i
d
e
d
b
y
p
u
l
s
e
c
h
a
s
e
e
x
p
e
r
i
m
e
n
t
s
(
i
.
e
.
,
c
e
l
l
l

a
b
S
e
P
r

i
n
D
e
i

a
r
t
o
f
t
h
e
d
i
s
c
u
s
s
i
o
n
w
i
t
h
a
r
e
g
u
l
a
t
o
r
y
a
g
e
n
c
y
a
t
a
p
r
e
I
N
D
c
o
n
f
e
r
e
n
c

e.

wi
th
Pe
liz
ae
us
M
er
zb
ac
he
r
di
se
as
e
w
as
ev
id
en
t
in
th
e
re
st
or
ati
on
of
th
e
tis
su
e
m
ic
ro
en
vi
ro
n
m
en
t,
as
in
di
ca
te
d
by
an
in
cr
ea
se
in
fr
ac
ti
on
al
an
is
ot
ro
py
an
d
re
du
ce

d
radia
l
diffu
sivit
y on
diffu
sion
MRI
132
.
The
restorati
on of
neur
oche
mica
l
tone,
as in
the
case
of
Parki
nson
s
disea
se,
can
be
deter
mine
d
relia
bly
and
quan
titati
vely
by
PET
using
the
dopa
mine
D2
recep
torbindi
ng
agent
,
raclo
pride
76,14
0

.
Redu
ction
of
raclo
pride
bindi
ng
corre
lates
to
the
prese
nce
of
endo
geno
us
dopa
mine

iv
e
A i
s m
ag
in
g
in
to
cli
ni
ca
l
tri
al
s
wi
ll
fu
rt
he
r
be
ne
fit
th
e
sa
fe
ty
as
se
ss
m
en
t
(e
.g.
,
s
m
all
bl
ee
ds
,
ed
e
m
a)
an
d
qu
ali
ty
co
nt
ro
l
(e
.g.
,
lo
ca
ti
on
of
ce

lls)
of
cell
deliv
ery.
The
host
infla
mma
-tory
respo
nse
in
patie
nts
can
be
moni
tored
using
11
CPK1
1195
PET,
whic
h
binds
to
the
perip
heral
benz
odiaz
epine
recep
tor
on
activ
ated
micr
oglia
146
,
as
well
as
ultra
small
SPIO
upta
ke
into
perip
heral
macr
ophag
es
detec
table
by
MRI
147
.
In
rode
nts,
an
endo
geno

o
p
m
e
n
t

u
s
A
l
I
n

o
f
n
o
v
e
l

B
o
x
5
C
o
n
s
i
d
e
r
a
t
i
o
n
s
f
o
r
t
h
e
d
e
v
e
l

c
e
l
l
t
r
a
c
k
i
n
g
a
g
e
n
t
s
B
e
f
o
r
e
u
n
d
e
rt
a
ki
n
g
t
h
e
d
e
v
el
o
p
m
e
n
t

o
Af
s

C
on
fir
m
th
at
th
e
la
be
lin
g
m
et
ho
d
is
a
m
en
ab
le
to
sc
al
eup
of
th
e
ce
ll
pr
od
uc
t
in
a
G
oo
d
M
an
uf
ac
tu
rin
g
Pr
ac

tic
e
fa
cil
ity
.
E
ns
ur
e
hi
gh
se
ns
iti

vity
of
det
ecti
on
by
ma
xim
izin
g
the
co
nc
ent
rati
on
of
lab
el
in
cell
s
(co
nsi
ste
nt
wit
h
saf
ety
)
an
d
mi
ni
miz
ing
the
nu
mb
ers
of
cell
s/v
ox
el
for
det
ecti
on
in
viv
o.

Ev
alu
at
e
wh
et
he
r
cel
lul
ar
dy
na
mi
cs
(c
ell

d
e
a
t
h
,
p
r
o
l
i
f
e
r
a
t
i
o
n
a
n
d
i
n
t
e
r
a
c
t
i
o
n
w
i
t
h
t
h
e
h
o
s
t
)
,
c
e
l
l
u
l
a
r
k
i
n
e
t
i

cs
(
m
ov
e
m
e
nt
of
ce
lls
in
th
e
b
o
dy
)
a
n
d
vo
lu
m
e
of
di
st
ri
b
uti
o
n
of
th
e
ce
lls
m
ay
af
fe
ct
ef
fic
ac
y.
St
riv
e
to
li
mi
t
tr
a
ns
fe
r
of
th
e
la
b
eli
n
g
a
g
e

nt
fro
m
th
er
ap
eu
tic
cel
ls
to
m
ac
ro
ph
ag
es
or
by
sta
nd
er
cel
ls.
De
vel
op
m
et
ho
ds
to
tur
n
off
th
e
sig
na
tur
e
of
th
e
lab
el
aft
er
en
gul
fm
en
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( co
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funding agencies to work together to solve this tractable problem. An alternative solution would be the development of novel agents that label only
live cells, disappearing quickly after cell death. In addition, most cell therapies would greatly benefit from novel ligands or probes with increased
imaging specificity to detect dynamic changes in the hetero-geneous, injured host environment. Understanding the role of the host environment will
be crucial to improving the efficacy of cell therapies.
Many results in animal models are not predictive of the human response. For example, therapeutic results in rodents do not mean that the same
number of cells/kg would be safe and efficacious in patients. Similarly, imaging results demonstrated in rodent models do not neces-sarily translate
into the clinic, primarily because of limitations in clinical scanner technologyspatial resolution or voxel size, instrumentation (i.e., radiofrequency
coils and magnetic field strength), patient motion and image acquisition times. Investigating teams must be aware that the translation of regenerative
therapies from bench to bedside is research, so they are advised to be patient.