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2Departm
ABSTRACT We demona migration cflow resistancsistance chokverify the migshow lower lywas confirme
KEYWORD INTRODUC Cell migratgenesis. Signi90% of cancehave been disdepends on xtherapy in reacially for the laries, and dev p38γ mRNof p38γ genescell can undelack of RhoCeffectively lement. To charies, we devisdemonstrated
DESIGN AN Figure 1 shvised chip. Chydrodynamithe migrationto the right-hent is generatthe channel isydimethlysiloglass substratThree masksheights for thcapture gap channel (10 µcated device gration resistaemulating thelaries [7]. Thfrom 6 µm tothe number ocated in the u
LY
Yu-Chih ChDept. of Electrment of Intern
T nstrated a singchannel with rce, the device ke points in thgration platfoymphatic metaed by the fabri
DS: Lymphatic
CTION tion is an essificant attentio
er-related mortscovered and
xenograft modal time [3]. Hlymph node invices that can
NA is overexprs degrades theergo [5]. It is b, the knockdo
ead to cellularracterize the i
sed a single cd the capability
ND FABRICAhows the scheCells are loadcally captured
n channels; chhand side, andted by diffusis observed aftoxane) layerste by the stans are used tohe channel reg
(20 µm heigµm height). Fand four diffance choke poe geometry of he width of o 30 µm. Thef choke point
upper part of F
YMPHATBY A SI
hen1, Steven rical Engineer
nal Medicine a
gle cell migratresistance choallows for po
he migration corm, we used pastasis in-vivoicated migratio
c Capillary Inv
ential processon has been patality [1]. Sevare actively i
dels, which reHence, there is
nvasion procen also be potenressed in sevee efficiency ofbelieved that
own cells (GKr displacemennvasion capab
cell migration y of tracing sin
ATION ematic diagramded by gravityd on the left-hemoattractant
d the concentraon [6]. Cell mter 24 hours. Ps were fabricndard fabricatio fabricate thgion (40 µm ght), and the
Figure 2 showferent designsoints (100 µmlymphatic or the choke po
e motility wass passed by c
Figure 2.
TIC CAPINGLE CG. Allen2, Zring and Compand Cellular aversity of Mic
tion chip whicoke points. Uositioning singchannels are ip38γ gene kn
o in a separateon chip.
vasion, Single
s in angiogeneaid to the mig
veral metastasiinvestigated [
equire a great s an unmet neess which is a ntially employeral types of cf lymph invasp38γ knockdo
KD) are unablent. The resultibility of MDAchip that con
ngle cells in th
m of the de-y flow and
hand side of ts are added ation gradi-
migration in PDMS (pol-cated on a ion process. he multiple height), the e migration
ws the fabri-s of the mi-
m length) for other capil-oints varies s scored by ells as indi-
ILLARYCELL MIGZhi-fen Wu2,mputer Scienceand Molecularchigan, Ann A
ch can emulatUsing a hydrodgle cells at theinvestigated t
nockdown MDe study. The re
e Cell, Cell Mi
esis, cancer mgration of cancis-suppressor [2]. Neverthelamount of tim
eed to developcritical step in
yed for therapecancer and helion in-vivo paown leads to
e to form longing unorganiz
A-MB-231 (brntains multiplhis lymphatic
Figure 1. Swith invasio
Y INVASIOGRATIO, Sofia D. M
e, University ofr Biology Progrbor, MI, USA
e cancer cell idynamic captue start of a migto characterizeDA-MB-231 (eduction of th
igration, Meta
metastasis, wocer cells sincegenes, which
less, the studyme and cost ap in-vitro devin the metastaseutic decision lps increase Rartly due to drubiquitination pseudopodia
zed cytoskeletreast cancer) cle migration r capillary inva
Schematic diaon resistance
ON ASSAON CHIP Merajver2,3 anof Michigan, Agram, and 3DeA
invasion in lyuring scheme gration channe the deformabreast cancer)e invasive abi
astasis
und healing, e cancer metas
may be poteny of metastasiand are difficuices which casis of cancer tmaking in the
Ras-induced carastically chann and degradaand thus enga
ton reduces thcells in a 3D mresistance choasion assay.
agram of the choke points
AY nd Euisik Y
Ann Arbor, MIept. of Epidem
ymphatic capilbased on the
nel. Different sation capabilit) cells, whichility in lympha
inflammationstases accountntial targets fois-related genult to adapt to
an emulate methrough the lye clinic [4]. ancer invasionnging the typeation of RhoCage in motilityhe efficiency model of lympoke points an
single cell m
Yoon1 I, USA miology, Uni-
llaries throughe difference insizes of the rety of cells. To
h are known toatic capillarie
n, and embryot for more thanor therapeuticses still mostlyo personalizedetastasis, espemphatic capil
n. Knockdowne of motion theC [5]. Due to ay that does noof cell move
phatic capillard successfully
migration chip
h n
e-o o s
o-n s, y d
e-l-
n e a
ot e-r-y
p
16th International Conference on Miniaturized Systems for Chemistry and Life Sciences
October 28 - November 1, 2012, Okinawa, Japan978-0-9798064-5-2/μTAS 2012/$20©12CBMS-0001 968
EXPERIME MDA-MB-penicillin/strewere stably scrambled ve[5]). Before cellsolution, whiLiquid heightflow, and thuthe cell soluti20% Fetal Boduce migratiobehavior is obby monitoring
RESULTS A First, we ochoke point mphologies of cells. F-actinwere able to felongated anstress fibers. knockdown, G Next, we oants without ng/mL hepatotion. After codependence o(Figure 5). Intractant since We observed migration chabut the motiliis obstructed b To verify thvelocity of Mµm x 10 µm) large, the veloresult confirmdue to a lack
Figure 4. Midifferent chem
NTAL -231 cells weeptomycin. p3transfected w
ector (SCR) ce
l loading, trypch are dilutedt difference beus cells are cion in the inleovine Serum (on. Then, the bserved after 2g every 30 min
AND DISCUSobserved cellmigration chanscrambled ve
n fibers are laform long psend migrated thDue to the la
GKD cells couobserved cell resistance ch
ocyte growth onfirming chemof cell motilityn this experim
HGF may alsthat SCR and
annel is wide ity of GKD ceby narrow chohe lower migr
MDA-MB-231is measured a
ocity of SCR ms the hypothof RhoC. The
igration distamoattractants
ere cultured in38γ knockdowwith short haells were the
psin/EDTA isd to 106 cellsetween the inlaptured hydro
et is replaced b(FBS) media ientire chip is 24 hours, andnutes.
SSION l migration bnnels. Figure ctor (SCR) ceabeled by redeudopodia pashrough the naack of RhoC uld only squeemigration di
hoke points, afactor (HGF
motaxis of My on the dime
ment, we used so regulate Rhd GKD cells h(30 µm x 10
ells significanoke points (6 ration efficien cells in the nas shown in Fcells is almos
hesis that the e actual distrib
ance of MDA-without chok
n RPMI with wn MDA-MBairpin RNA standard cell
s used to re-ss/mL and pipelets and outleodynamically.by serum-freeis applied to tput into an in
d the velocity o
behavior in th3 shows the
ells and p38γ d fluorescencest the choke parrow channeand other gloeze into the nastance for vaas shown in ) and 20% F
MDA-MB-231 nsions of mig20% FBS mehoC and confohave a simila
0 µm) and wintly diminisheµm x 10 µm).
ncy of p38γ knnarrowest choigure 6. Althost double that motility of G
bution of cells
-MB-231 cellke points.
10% FBS anB-231 (GKD)(shRNA), anline (Merajve
suspend the cetted into thets generates g. After 10 mie culture medithe other inletncubator. Migof cells is mea
he 6 µm resirepresentativeknockdown (e. Since SCRpoint, the cellsel by contractobal effects ofarrow channelrious chemoaFigure 4. BoBS induced mcells, we test
gration choke dia as the cheound the resular motility whthout choke p
es when the ch. nockdown celke point chan
ough the variaof GKD cells
GKD cells decfor various ch
ls for Figurand G20% F
nd 1% ) cells nd the er lab,
ells in e inlet. gravity inutes, ia, and t to in-gration asured
stance e mor-GKD)
R cells s were tion of f p38γ l. attract-oth 50 migra-ted the points
emoat-lts [8].
hen the points, hannel
lls, the nnel (6 ation is s. This creases hoke
Figed
Figbelepoinp38
re 5. Average GKD cells foFBS as the ch
gure 2. Micropd device and s
gure 3. MDA-Med by RFP) innts : (a) scra
8γ knockdown
number of cor various chemoattractant
photograph ofsize variation
channels.
MB-231 cells n the 6 µm xambled (SCR)(GKD) Cell.
choke points poke point dimt.
f the fabricat-of migration
(F-actin is lax 10 µm choke) cell and (b)
passed by SCRmensions with
-
a-e
b)
R h
969
point dimensichoke points. CONCLUSIO In this worlymphatic capplaced at the behavior of cashow that thepoint channelchip, and the ACKNOWL
This workChemical Genliance AwardUniversity ofFoundation, tand the Natio
REFERENC[1] D. Hanaha[2] A. N. Sho
lost in tra[3] N. Sethi a
Nature R[4] S. D. Nath[5] D.T. Rose
rajver, p3architect
[6] Y.-C. Checeeding o
[7] S.-Q. Zhoics in rab
[8] L. Jane, ATherapy,
CONTACT *Y.C. Chen, t
Figure 6. Athe migratioµm.
ions is depicteThe heteroge
ON rk, we implempillaries by adstart of a migancer cells in
ey had significl. Due to the ndifference bet
LEDGEMENTk was supportnomics at the
d from KAUSTf Michigan. Tthe Tempting
onal Institutes
CES an, and R. A. oushtari, R. Zanslation?, Naand Y. Kang,
Rev. Cancer, 1hanson, Insighenthal, H. Iyer38γ promotes ture, and a noven, X. Lou, P. of MicroTAS,
ong, Y.-D. Xu,bbit stomach, A.M. Tracey 6 (2), 173-17
tel: +1-734-56
Average velocon channel wi
ed in Figure 7eneous behavi
mented a singledjusting the chgration channea 3D model.
cantly reducednature of singltween motile
TS ted in part by Life Sciences
T. The cells arThe Merajver
Tables Orgaof Health.
Weinberg, HaZ. Szmulewitzature Rev. CliUnravelling t1, 735-748, 2
hts into the Mer, S. Escudero
breast cancevel leading edIngram, and 1409-1411, 2, Y.-F. ZhangWorld J Gastrand G.J. We
79, 2011
65-9976; yuch
ity of MDA-Mith choke poin
. Most cells por of cells in t
e cell migratiohoke point dimel. The novel As a proof of
d motility as cle cell resolutand immotile
the Thermo Fs Institute at thre cultured bygroup receive
anization, the
allmarks of caz and C. W. Rinical Oncologthe complexit011. echanisms of L
o, L. Bao, Z. Wer cell motilitydge behavior, E. Yoon, Sing011. , Y.-F. Zhangroenterol, 4(6
en, HGF and
hchen@umich
MB-231 cellsnts of 6 µm x
assed fewer ththe channel ca
on platform wmensions. Usin
design of resf concept, p38compared to sion, heterogencells can be f
Fisher Scientihe University
y the Merajveres funding froDebbie Stran
ancer: the nexRinker-Schaefgy, 8, 333-342ty of metastas
Lymph Node MWu, A.C. Venty and metastaCancer Res.,
gle cell Migra
, L.-S. Hai an6), 550-552, 19
rhoGTPases
h.edu
in 10
Figure cells) asious dim
han two chokean also be stud
with choke poing a hydrodynsistance chokeγ gene knockcrambled cellneous cell popfurther studied
ific Screeningof Michigan,
r laboratory atom the Breastnge-Browne In
xt generation,ffer, Metastas2, 2011. sis — molecul
Metastasis, Ctura, C.G. Kleasis through r71(20), 6338-
ation Chip Usi
nd F.-C. Tang, 998 in cancer ce
7. Distributios a function of
mensions.
e points and vdied in future
ints to emulatenamic capturee points allowdown MDA-Mls when migrapulations can d in the future
g Technology , and in part bt the Compreht Cancer Resenflammatory
Cell, 144, 646sis-suppressor
lar understand
Cancer, 98, 413eer, E.M. Arruregulation of R-6349, 2011.ing Hydrodyn
Three-dimens
ll motility, Cu
on of cells (SCof passed resis
very few cells work.
e cancer cell ie scheme, singws us to studyMB-231 cells ating in the nabe labeled an.
Grant under by Academic Ehensive Canceearch FoundatBreast Cance
6–674, 2011. r genes in clin
ding and targe
3–423, 2003 uda, K. GarikiRhoC GTPas
amic Cell Pos
sional structu
urrent Signal
CR and GKD Mstance choke p
passed all five
invasion in thegle cells can be the migrationwere tested to
arrowest choked traced in the
the Center foExcellence Aler Center at thetion, the Avoner Foundation
nical practice
eted therapies
ipati, S.D. Mee, cytoskeleta
sitioning, Pro
ure of lymphat
l Transduction
MDA-MB-231points for var
e
e e n o e e
or l-e n n,
e:
s,
e-al
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n
1 r-
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