Y1D

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

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or l-e n n,

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