CA2000254A1 - Stabilized compound of fgf-family growth factors and dextran sulfate and it a applications - Google Patents

Abstract

"Stabilized composition based on growth factors of the FGF family and dextran sulfate, and its applications" Invention by: BARRITAULT Denis CARUELLE Jean-Pierre COURTY José INSTITUTE OF DEVELOPMENT AND BIOTECHNOLOGICAL DEVELOPMENT This composition includes the combination of at least an active form of a growth factor of the FGF family and at least one dextran sulfate stabilizing the biological activity of FGF. It is applicable in the fields of medicine, surgery, cosmetology, biotechnology and cell biology, and more particularly, in the fields where growth factors can be used as therapeutic agents, in particular in healing and tissue regeneration, as well as cosmetic agents. No figure for the abstract.

Description

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; P ~ SIl '~ STAeILI ~ EI ~ A ~ / ~ 5E ~ E FACS ~ U ~: ~ E CROIS ~ ANCE ~ 1 ~
LQ l; '/ ~ MI LLE l: ~ F ~ T kE ~ EXTRANE ~ ULF ~ Al' ~, E 8E ~; ~ PPLICATI4t ~ J ~.
L ~ pre ~ ent ~ inventlon se! stop ~ t ~ ~ A ~
compo ~ ltlvn ~ to ~ b ~ ed '~ 1 mo ~ n ~ u ~ f ~ e ~ eur d ~
crols ~ nc ~ dc 1 ~ femllle FGI ~ and do d ~ xt: ~ l ~ aa ~ ul.fot ~, ~ ln ~ l c ~ u ~ ~ u, ~ ~ ppllcc ~ tior, ~ de cett0 compo ~ tlon z ~ to ~ ed ~ n3 ~ e ~! s domolne: 3 de l ~ m ~ decine, d ~ l ~ ch ~ rurlsio d ~ 1 ~ co ~ méto-lo ~ le, d ~ 1 ~ blc ~ technoloF ~ e and l ~ blo v ~ c ~ llul ~ lr ~ r and, more partl ~ llLère: ment, clQns lo ~ do ~ c .n ~ e ot ~ les ~ qCteL ~ rg l ~ J de CfOl5 ~ nce can be l ~ t ~ lls ~ com ~ ~ ~ g ~ -nts ~ th ~ rsp ~ u-tlque ~, not ~ nt er ~ clc ~ trla4tion and f ~ 5 ~ r ~ t ~ on ti ~ ul ~ ir ~, ~ in ~ i as like ~ ent8 cosm ~: lqu ~ -~ 4 ~ cover ~ rt ~ d ~ l ~ f ~ m1 llo ~ B factaur i ~ e cr ~ anç ~ des flbroblsste3 ~ courc ~ @ nt ds ~ 5n ~ P ~
l ~ l '~ br ~ Yiatlon (~ FGF ~ - Fibrobln ~ t Orowth 4ctor ~ a ~ t ~ f ~ lt ~
ulte des e ~ .ttld ~ de ~ ~ tlvlt ~ ble: ~ ~ glq ~ u ~ of ~ cte ~ r3 d ~
crol $ snnc ~ obtained don the leaps of n tr ~ rond nomorq ~
d ~ tis ~ u: s or ~ 'or ~ s ~ n ~ s ~ c ~ calf. hypovhj s ~, r ~ t ~ n ~. hunn6! u vitr ~ e, ehoroïd ~, lrl ~ ortil ~ e, re ~ n, ~ oi ~, pll ~ cents, or ~ ut ~ m. pl "oste ~ t ~ o ~ n ~ e, and). L ~ dlv ~ r ~ it ~
m ~ m, ~ dl ~ 3 tlss ~ s ~ studied ~ 3 4t d ~ sc ~ lul ~ tlmul ~ -sJ p ~ r ~: ~ 3: ~
t'e - :: tl ~ lJrs ~ 'nv ~ ro Ct ln ~ iYC ~, ~ ln ~ l que l ~ ~ r ~ dn ~ b d '. ~. ~ u1p ~ ~ ls rech ~ rche qu ~ have ~ nd ~ pen ~ mm ~ nt contsq ~ b ~
c ~ rnc ~ ti-on, bl 'l: ~ o). ~ m ~ nt ~ t ~ l' 1 ~ n ~ ti ~ n

2 compl ~ te to ce ~ fsctcurs, expllq ~ 2nt ll ~ a ~ ultltu ~ e de ;; no ~
at ~ 51gle ~ given ~ por ce ~ ~ it f ~ nlt ~ ~ utqur ~ for ~: ~
d ~ 31 ~ ner. It ~ ppar6 ~ t ~ ln ~ nten ~ lnt tl- f ~ on cl ~ lre qu ~ 3:
tou ~ s ce ~ ~ xtt ~ ait ~ cont L ~ 3nne! nt d ~! ~ faet ~ r ~ ll d ~ ~ rol ~ snnce do the f ~ mlll ~ of FQF3 and that cott ~ af ~ ll p ~ ut ~ be

3 ~ r ~ pr ~ nt ~ e por d ~ u: c br4n ~ he ~ 3 pr ~ nclpal ~ ~:
L ~ premi ~ r br ~ nch ~ t ~ d ~ C It- ~ ousl l ~ fi names de F&F bQsiq ~, de f ~ ct ~ ur de Cr ~ Oi ~ S ~ nCQ b oiqu ~ Ou d- typ ~ 2 a ~ n po ~ rl ~ h ~ p4rln ~ or ~ 3porln ~ ind ~ ~ O ~ c ~ th l ~ octoro (~ UCI: II), dG! f ~ ctel ~ rd ~ ~ rol ~ nc-3 çl ~ rv ~ du ~ rvotau or 3.5 ~ r ~ ln ~ lv ~ sd Crowth ~ actor II 5 ~ FX ~, d ~ ~ nct ~ 3ut ~ de crol ~ G ~ nce d ~ rlv ~ from o ~ ll or E; re-D ~ l ~ Ct ~ owth ~ tor I
,, . . . . . ... . . . . . . .

Z ~ S ~ l i '~ T' ~ 3 P ac te ~ lr c ~ ec roi ~ nc ~ d ~ 3 ~ tr ~ Cyt o ~ IX
A ~ F II, d ~ fnctel ~ r de croiS ~ nCe d4rlv ~ ~ u co ~ ei1 tc., DI o ~ u ~ lo deux ~ ab 4r.che cl ~ t om1 1 1 ~ 0 ~ t ~ d ~ crlt ~ sou ~ the names $ tle FCF ~ 1'd ~ or f ~ ctc \ Jr 'j af f ln po ~ lr hepar1ne do ~ ype I Ha ~; FI, f ~ ctour d ~ rlv ~ du cerve ~ u or B ~ OF X, fc ~ ctour de crolu ~ nl; ~ d ~ rive d ~ 1 'o ~ ll or E ~) CI; II, ~ ef ~ ctollr d ~ crol ~ s: 3rln ~ lo I ~ $ trocytc ~ u ACF I, f ~ ct ~ ur de cro ~ nco d ~ rlv ~ u cart ~ lnEse ~ CDCF I), etc. A tlt ~ er ~ r ~ nc ~ b bllo ~ r ~ phiqu ~
concerned ~ t the ~ cruiser ~ 3 ~ nce, n ~ lt ~ r4 y RLobb, d ~ o ~ 3 ~ An ~ lytic ~ l Bloch ~ mLo .ry ~ a4 ~ Ig ~> o, and J. Courty ~ t coll, dc ~ n ~ ~ h ~ m ~ Blophy ~. Re ~.
Comtnu ~ 10 ~ -10 ~ ~ 98G ~.
These ~ u ~ taur ~ ort therefore and ~ norn ~ o, or solon the ~ yp ~ d ~ cell ~ clble $ utili ~ s ~ f ~ ct ur3 cl ~ ~ oio ~ nc: ~
dc: s fiDrobl ~ 3t ~ trocyt ~ s ou ~ llule endoth ~, eve ~
lq3 ~ lgll93 r ~ pe ~ tlvem ~ nt FCF, AC ~, ECO ~, oo ~ t 5elon lo . ~ OI, II ~ C ~ 3 d 'o ~ ce fuct ~ ur ~: ~ t extrQit ~ p ~ re ~ compl ~, f ~ ct ~ ur d ~
croi ~ ce d ~ r ~ v0 du cerv ~ u, t ~ d ~ 1 ~ r ~ ln ~ s ~ ud ~: 3 ) yeu ~ o ~ l-til ~ 3, h ~ p ~ tocyt ~ in cu ture, ~ lc;
s1 ~ 1eo r ~ _: 3p ~ ctlv6 ~ ment B ~ ..F, RClCF, Ft ~ P ', C3GE ~, HI ~ GF, ~: t ~. ).
s ~ lt still ~ ~ ulon un ~ proprl ~ t ~ b ~ o ~ him c ~ u ~ ou b ~ olo ~; Lque (factor, ~ ~ c ~ ol ~ u ~ ce ~ f ~ p ~ ulr 1I h porl ~ 0 ~ ~ u î nct ~ r tumor ~ l ~ n ~; lo ~ nlq ~ R ~ TI ~ ~ au; c prin ~: lp ~ r, ,: 'i- ~ br ~ ncl-e ~ d ~ l ~ t ~ emllle l ~ tAllt ~ lor ~ nomn ~ lon co ~ æl ~ lo ~:
pr ~ c ~ s or 3ulvls d ~ l ~ mentis ~ n ~ 4c1dl ~ ou ~ b ~ iquQ--, or: lo m ~ ntlon l ~ typ ~ - or ntyps II ~.
C '~: 3t ~ n ~ llv ~ nt l ~ t ~ ~ lt ~ bo ~ oE ~ u ~ ~ ur de ~
C ~ llu ~ n cultl.lre qu ~ ee ~ f ~ ct ~ ur ~ on pll ~ tr ~ ap ~ rlf ~
L ~ prem1 ~ r ~ cor ~ ct ~ rLot ~ qu ~ phy ~ l ~ o- hlmique ~ ~ poldo n ~ ol ~ eulolre et point l ~ o ~ lectr ~ qu ~) orlt ~ t ~ 6 pub ~ sp ~ r Co ~ ipocl ~ rowlcz,, r, Biol. Chem. ~ EQ. 2 ~ lY C147Y ~ po ~ Jr ln ~ orm ~ 3 b ~ 3Lqu, 3 ,. ~ tp ~ r ~ i ~ rrlt ~ ult et al., J. N ~ 3uroscl. I9L 47 ~ 0 (19 ~ z> for lcl form ~ ~ c1de.
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~ - Zil ~ r ~ 5 ~ l l ~ o purificntlon ~ homog ~ n ~ lt ~ ~ e ~ d ~ UK fqrm ~ u r ~ l ~ permi ~ ~ 'ét ~ bllr l ~ urs ~ tructure0 prlmolre ~; E. ~ ch r :, ~ t.-oll., Pro ~. Notl. f \ c ~ d. Sc ~. I.JS, ~ ~ !! S07 ~ 15B ~), p ~ u ~
1 ~ ferln ~ n 4 ~ 1qu ~ a, e ~ Oirnenez GÇ ~ SO and C411., 5clonc ~
l ~ 85 ~ l'385), po ~ r the form ~ c1 ~ 3. L ~ lsol ~ men ~ di ~ ls d ~ x forrn ~ t ~ ~ r, ~ ndement felvorl ~! por la dlSeouv ~ cte ~ 'un ~
forte nfflnlt ~ de C06 ~ fscteur ~ pour 1 '. h pel ~ ln ~ ~ t 1 'ut 11 i ~ aS lon subsé ~ uent ed' un ~ chrc ~ at ~ r ~ phl ~ d '~ f lnl t Gur h ~ parine lrnlmoblll ~ e (Shlng et al, Scl ~ nc ~,. ~, 1b 1296 ~ 9 ~ 1984)).
In v ~ tro, le33 FOF ~ ~ ont cnp ~ be ~ ~ e ~ ln prol ~ f ~ rntion ~ t 1 ~ dlff ~ r ~ nci ~ tlon d 'u gr4nd nonlbr ~ ~ e celll ~ lo ~ 3 prov ~ R ~ nt ds tl ~ sus ~ te ~ p ~ c ~: ~ dlff ~ rent ~. ~ n c ~ tcr4 cells ~ flbrobl ~ .stlques, en oth ~
15 ~ plt ~ la. ~ Sk ~ r ~ tlno ~ yte3, ehondr ~ y ~ o, myo ~ l ~
~ stro ~ 5 ~ tq3, ~ t ~ n ~ n ~ lera ~ g ~ n 10 ~ Ole d ~ FGr :: ~
d, 3n.s 1. ~ neuron ~ survival. In v ~ ~ rc ?. 1 ~ property ~ D
ncurotroph ~ ue3, anE5lo ~ enique: P ~ t Si ~ loonte ~ dr ~ FC; Fs ol ~ t c ~ n; qnt ~ t ~ d ~ shown ~ - ~ sd ~ r \ s of NUM ~ Ix ~ yot ~ m ~
XO L ~ r ~ l ~, dqn ~ la Yttlmlll4tlon ~ 1 ~ cl ~ oL ~ 9d ~ d.
1 ~ pid ~ trms, of an EXTRE ~ it ~ qu ~ ux de r ~ tl ~ p4rtl ~ m ~ nt purifl, ~ and therefore contain FGF comm0 ~ lnclpe nctlf, e- ~ t indlqu ~ d ~ ns 1 ~ br ~ vqt fr ~ n ~ 3 n ~ 2 4 ~, OO ~. L ~ r ~ e C ~ ~
same ~ :: very 5U ~ cicutrls ~ tion of ~ pl h ~ llum d ~ c ~ ln * ~ ~ t ~ 5 indl ~ ué à ~ n ~ le brev ~ t ~ m ~ ric ~ ln n '4 ~ ~ 7 435. ~ pul ~ ic ~
of no ~ rellx br ~ v ~ ts were ~ d ~ llvr ~ d ~ norn ~ r ~ u ~ ce ~ -publlc ~ tiono s ~ lentl ~ lque ~ ~ 3nt poru, ~ u ~ r ~ nt lo r ~ l ~ d ~: 3 I ~ Crs purlflé ~; t ~ n ~ 3 la r ~ nérc ~ lon ~ t 1 ~ le ~ eh ~ rl3 ~ ion of 1 ~:.
p * ~ u, dl_ $ V ~ iS ~ x, de, sr, erF ~, d2 ~ 4 09, d- ~ muscls: ~, ctc.
Most ~ rt: 1 ~ c ~; 3 trav ~ ux r ~ ult ~ nt d * 4 tron ~ pa ~ tlon de ~
obsarvot ian: ~ o ~ t3ur outfits ~ sc ~ lluli ~ s ~ n eulture ~ u; ~ m ~ m ~ 4 1 U l ~ v ~ ~ c7, / ~ ln ~ l, 1 '~ ppl ~ ~ ation ~ op ~ quo ~ ~ 3F ~ ul or 1 'utlli. ~ Atlon d ~ polyln ~ r ~, t' ~ pon ~, o ~ 1A OR ~ e pl ~ mp ~: ~
, 35 p ~ rmett ~ nt l ~ nr ~ larg ~ t de3 FGF ~, ~ Ut lnduiro urla n ~ oan ~ rl o $ ~ n ~ se ~ t url ~ r ~ n4r ~ t lon n ~ ~ U ~ or cut ~ n ~ e.

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~ hang, 1 'nppllcot lon topl ~ o from FGI: ocid ~ o ~
l) ~, l ~ u ~ olt: ~ o ~ nt ~ tr- ~ lt ~ r ~ t'Lve pol obter ~ lr 1 ~ cffet ~
mQ: clm ~ l.Jx. C ~ cl 6: 3t probably dC ~ 4 u ~ e, ln ~ obll ~ t ~
ther ~ n ~ qll. ~ of 1 ~ rnol ~ c ~ le, a proteolyr p ~ r dlæ3 in ~ ym ~ 4.
! ~ ~ lnsl qul ~ IJn ~ lnteract ~ n ~ ntre le ~ FO c et 1 ~ lyco4mlno-zlycones, col ~ une 1 'h ~ p ~ r ~ ae sulfate ou p ot ~ oh ~ r ~ n ~
sulf ~ t ~, de9 meml ~ relne ~ eellulslri ~ a or ~ ~ mambr ~ ne ~ 3 balcQle ~, e ~ r ~ fn ~ nt une lmmoblll ~ tlan de ~ a F ~ F ~ c.uvant emp ~ cher leur acc ~ ~ ux r ~ eepteur ~ e ~ llul ~ ro ~. At c ~ propo8, a ~ ~ b4ence ~ e 3tlm ~ Jlotlon de l ~ cl ~ z ~ tri ~ tlon p ~ r ~ FGF d ~ p2, ~ 1e ~
profon ~ -3 ~ ~ t ~ r ~ pport * e (J. D ~ rm ~ tol. 5urg. Oncol. t ;, 6 l 7-~ 25 ~ 19 ~ 6 ~) lor ~ that the Fap s ~ ul ~ t elpp ~ qu ~ a m ~ nlbrq toplqu ~, ~ when ur.e pr ~ l ~ f ~ r ~ tton fl ~ o ~ lu3t ~ q ~ ~ t ~ yobl4s ~, lqu6 ~ t obtainedR lor ~ qu ~ de ~ ep n ~ eonst ~ killed ~
1! ~ D '~ lc ~ ol polyvlnylique l ~ nbib ~ ~ e ~ GF ~ ot oppllqu ~ sou ~ 1 paau (J. ~ lol., 1 ~ 0, 12 ~ 9 C1985).
We ec ~ nst ~ te p ~ r ~ llleure qu ~ ~ FCF, ~ u ~ ol b ~ n in ~ form ~ cl ~ F ~; F) ~ u ~ ~ OU5 ~ 9 ~ elm bz ~; lque ~
~ eu 1 'we keep ~ 3 h ~ bltuelle ~ m ~ nt ~ -80', p ~ r ~ d- ~ J ~;
ctivite lor ~ que 11 e3t ~ m ~ né t. -20 '~ co ~ ~ templ ~ r ~ tur- ~
bi.3rlr, ~.
~ 1 ~ pp ~ r ~ it therefore 1 ~ c ~ 31t ~ d ~ 3t ~ blll; ~ 3r 1 f ~ cteul- ~ ds cro ~ s ~ 4nc ~ F ~ 3 ~ for p ~ uvo ~ ~ nvls ~ ser r ~ mont their ;, ~ pplle ~ tiorl: s. For the ~ ppllent ~ no ~ n vf ~ o L'5 notllmtnent, 1 s ~ r ~ lt avt ~ nt ~ eu ;; qu ~ ~ odG ~ d ~ ~ t ~ t lon the ulese will allow a relc of P; ~, Cl3 d ~ rn ~ qr ~ even ~ nt un aE ~ 3nt mito ~; ~ ne loco.l, r ~ ntr ~ nt d4n ~ le8 proces ~ on ~ turq ~ l ~ d ~ r ~ n ~ r ~ tion c ~ ll lult ~.
t ~ 1 'hol ~ te ~ ctu ~, compte t ~ ~ ud ~ 1' lmpc ~ rt ~ nc ~
d ~ G fnct ~ urs de cro1 ~ s ~ n ~ e dor ~ s lo ~ do ~ iln- ~ pr ~ c ~ eé3, on ~ lt q ~ c baAucoup d ~ t ~ nt ~ lb ~ ve: ~ ~ ont ~ f ~ ctu ~ a pour 3t ~ blll ~ rl ~ CFs, m ~ eonn ~ l ~ ~ nc ~ 3 la D ~ ent ~ 3, there ~ pQs ~ ncoro eLI ~ c ~ o ~ r dl3 propo ~ tion conclude ~ t ~ ~ t 4 ~ t1 ~ fo1 ~ nnt ~ for ~ t ~ b1114 ~ rl ~ o ~ for : 3'j que 113 con ~ e ~ -v ~ nt l ~ ur ~ ct1vitS r.on ~ - L ~ nt ~ n ~ ro, mol ~ 3 , s ~ m, ~ nt ln vi vo.

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t lon ~ u pr. ~ b ~ oY. ~, cor ~ t ~ nt ~ eoc ~, on un ~ tr, :) n.
., ~ ulr ~ J ~. ~ aux F ~ i ~ s, 1 ~ con ~ itlc ~ nr ~ ~ r ~ t ~ ~ t ~ nt etclbla ~ u ~ to ~ M ~ mæ ~ temp ~ r ~ tur ~ nbl ~ nte.
L ~ pr ~ nt ~ lnventlon ~ ~ onc pqu ~ obJ ~ t a ~ o ~ npo ~ lti ~ n co ~ npren ~ nt 1 '~ 3. ~ 0clot ~ L nd' ~ u molns a rorme ~ ctlv ~ ~ 'un ~ cteur d ~ c ~ ol ~ 4n ~ d ~ la ~ ml ~ l ~ FGF ~ t d '31- moin. ~ un ~ extr ~ n ~ eul r ate st ~ blll orlt 1' ~ Ctlv ~ t ~
blelo ~; ic of FC; F.
1 ~) Le ~ de: ct r ~ n ~ s 6ul f ~ tes ut ill ~ ~ have d ~ 3xt r ~ n ~ e . ~ ulfat ~ .P t, y: lr4 ~ 0lub ~, $ n401uble! ~ cn i ~ lou o ~ u ~ ux olJ r ~ -l3 In- '~ 3 IJ ~
L ~ tr ~ r, ~; t; ulf ~ te4 ~ ~ lub oer qu-t 1 'oo can ~ It ~ llsal- p4ur 1 ~ mi: 3 ~ en 02uvre ~ e 1 '~ Yon ~ lon, ~ nt do ~
1 ~ p, r, ~ uits ~ olJr ~ nt ~, or ~ ~ a r ~ i-r4:; ~ nt ~ p ~ r 1 ~ forrr, ul ~:
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d ~ n ~ u * lls u ~ h trols ~ roupe: ~ hydro yl ~ du n ~ Ot ~ r ~ onom. ~ re s ~ "lt ~ t 1 t U \ ~! ~; p ~ r cle. ,, ~ roup ~ -0-503 ~ forme ~ ld ~) 0 .5 - C ~ - .; f ~ (f ~ r ~; le ~. Le :; produl ~ s oo plus ~ u ~., Ont d ~. ~ p ~ lt. ~ l ~ $ sou ~ 3 forn ~ ad ~: ~ el: sd ~ ~ odl ~
L ~ 3: 3 products pr ~ f ~ r ~ p ~ Uv ~ n ~ tr ~ o polyn ~ br ~ 3 ~ e ~ n ~, tA M ~ cul ~ lr ~ cQmpr ~ se ~ nt ~ ^ e 1 ~ 000 ~ ~ 00 I ~ O, nt. ~ ntnt nt, 2r ~ tr ~. ~ 000 and 5VO 00 ~, tol3 ql, l ~ 1 'on p ~ ut e.' ~ n . ~) præc ~ kr ~ up ~ d ~ 1 ~ Socl ~ t ~ ~ ICMA ~ R t` ~ 07 ~, Dl ~ 39 ~, ~ OC ~ 7 l 40, ~ 3 ~ 06 of the cPt e ~ u ~ l9 ~ S ~).
L ~ 3 ~ xtran ~ ulfAto ~ lnfi ~ c ~ l bla ~ 40nt ~, e d ~: ~ tranæ ~ l3cul ~ t0 ~ r ~ tl ~ ul ~ e according to 13i ~ proc ~ d ~! ~ h4bltu ~ 13 ~ e ~ t ~ 3., ~ t, i ~ n, cn portleull ~ r ~ ld ~ '~ pLchlol ~ hydrlnk,; ~ r ~ 5.

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Colr, me ex ~ pl ~ cl- ~ e ~ E: d ~ xtrnr ~ ~ ~ ulf ~ to3 ~ on p ~ l ~ t clt ~ rc, ~ lui commercl ~ p ~ r 1 ~ ScelA 6 51 ~ oU ~ 1 r ~ r r. ~ r ~ tal ~ u ~ 19. '' tt ~ np ~ Ut ~ 41 ~ nt o ~ e ~
tf ~ l: ad ~ 3; ~ tr ~ nH ~ ~ e1 on lf ~ p ~ otc ~ colel d ~ rtp ~ Mll ~ t ld. ~ J. ~ t cctll, An ~ l. Blt.?ch8~n. .LQ. ~ -: 304 ~ 0).
~ \ u ~ ln t ~ eompa ~ iltlon ~ on 1 'lnv ~ ntl ~ n, 1 ~ .3 tr ct r \ t ~ lJ lf 4 t es hy P ro ~ olu ~ on t ~ nts ~ un ~
~ ollc ~ ntr ~ tlc? n dta ~ FO m, ~ p ~ rm ~ 1111tro d ~ m ~ l4r "7.4 n ~ 3l. L ~ ~ nc, ¢ rltrat1on ~ o ~ ~ xtr4r ~; t ~ ulf ~ tfa ~ 3 ln ~ all bl.
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1. ~ eompo ~ ltlon s ~ lon 1 '~ nvent on contlenS p ~ r aill ~ ur. ~ ~ vanto ~ eusernellt des qu ~ ntlt ~ a; ~ fsct ~ true ~ O, ~ 1 rg 3 ~ 0 Jl ~ t '~ u nlolns une forrn ~ ~ ctlve do F ~ p ~ r mlllllltre ~ u gr ~ mme de mblan ~; e final ~ F ~ dextr ~ no ~ fAt ~.
L ~ d ~ xtr ~ no ~ oul ~ e ~ te3 ut ~ ~ pour 1 ~ ml5c en o ~ uvro ~ prb ~ nt ~ lnv ~ nt ~ on ~ er ~ en ~ d ~ ~ upport b 1 ~
ml ~ r ~ tlon ~ t QU ropouplem ~ nt cel ~ ul ~ lr ~ They can ~ nt ~ tr ~:
lncorpor ~ s dan ~ tout ~ s form ~ do p ~ n ~ o ~ nt ~, or ~ ncorr on fll ~ of ~ uture used ~ s in chlrurE ~, d ~ me ~ nlbr ~ que 1 1 ~ fact ~ ur ~ 56aoci ~ ~ c ~ dextrcn ~ ~ ulf4t ~ o ~ f ~ ct ~ ur tout : only pui ~ lf ~ u ~ r Yer ~ los ti ~ u ~ ~ l 1 L ~ 3 ~ cteur: sd ~ c ~ olss ~ ncQ F ~ utl ~ d4n :; cett13 invention p ~ uv ~ nt be d ~ prodult4 n ~ rolo, ~ s prodult3 u :; d ~:; ynth ~ e ~ chl ~ iqlle. ~ ou dc3 pro ~ Lt ~ obt ~ nl ~ 3 p4r ~ e;: ~
I !; techniq ~ es ds recombln ~ l ~ on ~ tlq ~ lo. It ~ can ~ u ~ tr ~
to ol ~ lgine hum ~ lne or to come from A ~ tre3 ~ rn ~ lfere ~ t no n ~ m ~ lf ~ re: ~ s. I ln ~ t ~ en ~ r ~ ler ~ ent p ~ ~ e ~ aslr ~ d 'ut lll: 13r 1 ~ t '~ ct ~ r ~ utologu ~. In ~ f fet, 1 ~ hol ~ ~~ conse ~
v ~ tlon of 1 ~ ~ tructllrç of ~ 3 F ~ F ~; ~ u C ~ UF of f ~ olutlon, n! ~ 1 q, ~ Q ~ ~ t ~ lde ~ ur le! ~ prop ~ t ~ a ~ n ~ lero et in ~
ln, ~ lqu ~ l ~ t that lL y 'p ~ d ~ e sp ~ clflclt cl' ~ p ~ c ~ for the;. ~ ~
dc ~ for ~ lle3 blo ~ ctive ~ da: 3 ~ ril ~ E ~ l ~ nt, orl utlll ~ a 1 ~ 3 ~, t` ~ oua ~ for ~ n ~ purlfi ~ e ~ el ~ e, or ~ o ~ 30 ~ ~ orm.3 baalq ~ le, or a m ~ l4n ~; e dQ: 3 d ~ lux form ~, c I ~ 2neoro un ~ rl ~ 5 ~ U
; '~ un ~ n ~ loé5u ~ u mo ~ n ~ 1' un61 do ~ a ~ fo ~ ~, L ~ ropport ent rc ~ s PCF ~ 4clde and ~ n ~ l ~ uo d ~ n ~ ~, form ~ n ~ p ~ l ~ t Atr ~ d ~ n 'lmporto quQlle v ~ laur ~ ntt ~ 1 ~ daux for ~ e ~ pur ~, Dlff ~ ront ~ proc ~ d ~ 7 do purlf'l ~ lon pau ~ nt ~ tre utll ~ s ~ s for l ~ lar this ~ dcux f ~ rm ~ a ~ ad ~ CJP ~ p ~ rtlr d ~. -oour ~ n ~ turell ~ fi Cr ~ tlno. c ~ rv ~ u, h ~ ~ elphy ~, plqlc-3nt4, r ~ in, ~ tc. >, c ~ proc ~ t ~ nt eonn ~ 4 t. 4y ~ nt 4t ~ described.
by dc r ~ ombrollx ~ t0ur ~. LQ m ~ thod ~ pr ~ r2 ~ tc ~ llo déçrlto pe7r ~; or ~ ty ~ t ~ oll. d ~ ns; (~ Bloc1r ~ G7. Blophyiil, R ~ 3.
Commun.,, ~, 102-lOB ~ lg ~ m C ~ tto r ~ thod ~ compr ~ l7d un ~
~ 5 ~ topc ~ on ~ 3l ~ t ~ nt 0. trc ~ lt ~ r, pen ~ 4nt ~ n night, do l ~ xtr ~ lt t i61al.1 ~, 7n ~ pr ~: 3t7nc ~ d '~ elt ~ 7 ~ 1 ~ c ~ tL ~ ~ ~ p ~ t 3, o ~ ccl ~ lan ~

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~ n ~ l tout rl- ~ qu ~ d ~ ~ ontomlntlti ~ n vlral ~, ~ t ~ ~ ondulra IJne p ~ o-iflc ~ tion p ~ l ~ chrom ~ to ~ r ~ phie ~ ur h ~ 3rlne lmmob ~ e p ~ rlne ~ eph ~ ro. ~ HAR ~ 1ACI ~. Loo c ~ ix f ~ rm ~ oeLde ~ t b ~ slque d ~ FCF pe ~ vent Qln ~ l être 1 ~ o7 ~ q ~ ~ tf ~ par ~ 4s ~ nvec le. ~ 4utres prott ~ lnl ~ 8 OU lnd ~ vlduell ~ men, e3ve ~ un ~ b ~ r ~ d ~
plJ ~ and ~ suffl! 3 ~ lnt pour atre d ~ b4rras3 ~ e ~ o ~ u ~ ntlt ~ 5 *
~ lgn- ~ lcatlv ~ .sd 'autr ~ mdt ~ rl ~ l ~ con ~ ~ nelnt ~.
~ es ~ nalo ~ ue ~ ~ ynth ~ tf que ~ ~ u ~ 0 ~ p ~ uvent att ~ e obtained pnr synth ~ chlmiqu ~ or pnr ds t ~ chnlquo ~
cll333 ~ c3u ¢: ~ d '~ nz ~ nl2rle ~ n ~ tlqu3, t ~ ll 1' oxpr2! 4 ~ 0n: le g ~ r, ea synth ~ tiq ~ es or l ~ pr ~ slon ~ n ~ o mocl ~ fl ~ pcr t ~ z3-3 diri; ~ d ~ lte ~ ~ p ~ lq ~
To ~ lt ~ a 1 ~ formQ: ~ de FGF obt- u ~ nd '~ xtr ~ lt a tl ~ ul4irr ~ p ~ r in ~ eni ~ rl ~ nétlq ~ l ~ or p ~ r synth ~
cnlrnlque, qul ell ~: ~ 41ent or not ur ~ e ~ tr cture modlflée, ~ lr) sl qu ~ le ~ fr ~ gmenta en d ~ r ~ t, p ~ v ~ nt ~ ro ut11s ~ e ~
~ or- r ~ '~ aerYe que elle3 4 ~ o ~ ent ~ ctlve; l ~ bl log ~ uement - un te. ~ t ln vi ~ ro cl ~; nm ~ the one pr ~ po ~ cl-clp ~ p ~ w4nt etr ~
utlli ~, 2 ~ 3 as ~ ti on! I ~ of FOFc t ~ t 4 cl ~ x ~ r ~ 3 ~ s ~ .J lf ~ t d ~ nvellt ~ on p ~ u ~ nt ~ tne ~ ncorpor ~ e ~ dsn ~ d ~ Uppvrt ~
phy ~ 1o1o; 31 ~ uem ~ nt ~ c- ~ ptab1e ~ p ~ ur d ~ ppllc ~ t ~ on ~ ln v ~ v ~, pl1c ~ t1on ~ st une elppl ~ catlon loc41, p ~ vo1a e ;; tenno ~ ur 1 ~ pa ~ u or 1 ~ co ~ e, not ~ mm ~ nt pO ~ I ~ ~ cc ~ l ~ r ~ r 1 clcc ~ trl ~ s ~ t ~ on a ~ pltlie ~ t 4m ~} 110rsr ~ qu411t ~ do cett- ~
~ lcatri. ~ tion, or ~ ncc ~ re p ~ r v01 ~ 2 lnt ~ r ~ l ~ ur 1 ~ t or ~ n ~ 3 du oorp6 ~, p ~ r ~ x ~ mple d ~ no 1 ~ t ~ ltem ~ nt d ~ 1 'uLc ~ ro ds 1 '~ stom ~ ç. L ~ l ~ s ~ type ~ l! Tl ~ ud 'c ~ ppl lc ~ tlon, l ~ a coml ~ o ~ 1tion3 Compr ~ n ~ nt 1 '~ a ~ ocla ~ on ~ ~ F ~ F ~ ~ td ~
3 (~ d ~:; trnn ~ a oul ~ a ~ Q ~ ~ eront dlf f ~ r ~ nt ~.. 1 ~ n ~ lt ~ ro ~ d ~
3upport ~ p-3ut ~ trQ c ~ ctr ~ ont v ~ rl ~ bl at d4 ~ and ~ onc du typa at llcu d 'uppl ~ ccltion, ~ t prut pt ~ ~ Idr ~ la forln ~ Jn lit ~ uldl ~, a / i ~, '9s4 ~ un ~ cr ~ me, d'~ ~ ~ ol, ~ n tl ~ su ~ II) p ~ n ~ comn ~ o un ~ ~ o ~ o, tull ~ ~ oci ~ t ~ on: 3 ~ G ~ '3-d ~ xtr ~ neo ~ ulfnt ~ e ~ lnvention p ~ R ~ V ~ ~ ~ gal ~ n ~; ~ very ~
lncorpor ~ e ~ ~ un ~ tln ~ eon ~ tlSu ~ ~ r a collos ~ n ~, qu ~ l ., -. . . ~ - ~. ::.,,:
:.

z ~ z ~

that ~ n solk the typ ~, or ~ ncore, like Is ~ i ~ t Ci-d @ ~ 3, recou ~ n lr de ~ rlls de ~ utur ~
~ e3 forll ~ ulQtlons ~ qu ~ u, soR ~ '$ nv ~ rltlon 1; eront o ~ nt ~ eu: ~ erl ~ nt d ~ s conp ~ sitlon ~ conten rt d ~ t ~ mpons and salts allowing m ~ int ~ nlr 1 ~ m ~ l4ng ~ ~ u vol ~ ln ~ 'du pl ~
physlolog1 qu ~ et ~ u volslnc ~ de 1 ~ for q ~ lon ~ que ~
phy31c> 10s: 1 ~ eu ~ olt pH of 6, ~ t ~ 7, 4, ~ t foree lonlq, ue ~ O, 1 ~ 0.2M). We can ~ Joust ~ compolsl ~ OA dl ~ 8 ~ ddlt lf8, comrn ~ ~ e ~ nn ~ th ~ lqu ~ s loc ~ ux ~ p ~ r ~ x ~ pl ~ a lldoc ~ Ino) ~
d ~ o 6ub ~ 3t ~ nce b ~ ct4ric: ~ st ~ tick ~ ~ by e ~ pl ~, do ~ 6el ~ s de s ~ dlum, d 'l ~ rsent, 1QU ~ S d ~ rlY ~ a ~ o ~ t ~ Ol ~ tu ~ f4 ~ 1e3 ~ 1n ~ e ~ 3 produl ~ t ~ epti ~ nts, ~ comm ~ B ~ tad ~ n ~, deJ ~ n ~ lblotlq ~ t ~;
~ p ~ rc:; ~ m.ple, 1 ~ streptomyclne), ~ e ~ pr t ~ n ~ ~ t ~ que ~ ~ p ~ r ex ~ ple, ln ~ rum albumin ~ or l ~ flbron ctin ~, as well as from t coll ~ n ~ ta hydr ~ solubl ~.
A tall ~ cornposltion ~ qu ~ us ~ r ~ nf ~ rm ~ nt ~ u mo ~ rls a f ~ cteur ~ CF ~ ssocl ~ 9 a ~ l moln ~ a ~ tr ~ n ~ 6ulfate ~. -hy ~ ro ~ vluble or insoluble, p ~ ut ~ tr0 u ~ ll ~ o dlr ~ oct ~ n ~ qnt 0n solutl ~ n, 6tn 41 ~ ro5tol, ell onction, en lo lon, ~ n poud- ~ ou ~ n 591, OR ~ el ~ vlr d ~ ~; olution pour ~ tr ~ ln lus ~ don ~ urt:; upp ~ rt ph ~ r ~ nacel. ~ tlqu ~ ment or co: ~; rn ~ tl4u ~ 3m3nt. ac e ~ ptoble, t ~ l ~ tl '4n ~ 1 apple ~ ds, a ~ crbme, a ~ el de re ~ no ~ a ~ 1 of a ~ ~
aut r ~ n ~ t ur ~, or 1 ~ t lne or 1 ~ ~ regn ~ r un ~ e ~ olla ~ n ~, un ~ 'unq ~ utr ~ n ~ tUr ~, RI d' ~ other ~ ul ~ qt ~ n ~
~ 5 fol34nt ~ on ~ sponge tion, a pQna ~ e ~ m ~ nt a ~ uppore d ~
overlay or biom ~ at ~ key ~ vtlnt fn ~ orlsor dir ~ ct ~ m ~ nt or ~ ndlre ~ ement 1 ~ r ~ p ~ rA ~ ~ on tlo4iulalr2 tp ~ r ax ~ mpl ~, a fll of ~ ut ~ ro chlrur ~ Slc ~ l) or ~ l; ro ~ ppllqu ~
~ ur d0 ~ flbr ~ na ~ ur ~ lleo oU rynth ~ tlq ~ ~.
"~ O For d6! S oppli ~ t ~ or ~ r 1 ~ I ~ U, lo ~ ba ~ de cr ~ m ~ or lotlon ~ used ~ trcldltSonnq Llo ~ r3t p ~ uvent ~ tr ~
utlli4i6 ~ s, Comme 1 ~ l ~ nolina (Silv ~ d ~ n ~ ~ M ~ arlon plu: ~
by ~ t lcull ~ rl ~ mant pcur le! .5 bru ~, ~ sJ Aquql ~ or Puk ~ 2 L ~ bor4tor1 ~ 0 ~; o ~ th Norw41k, C: onn ~ ctleu, l ~ qu ~
~ Gold ~ l)). Leo ~ rbm ~: j peuv ~ nt cont ~ n ~ 0 prot ~ 1no ~ ~ p ~ r ~: ~ elnplc, ~ oll ~ n ~ tins2, ~ te. ).

,.

`` `z ~ 2S4 -.
L ~ p ~ ns ~ m ~ nt. can be from pono ~ mento cl ~
toxtll ~ .i, tls ~ u ~. ~ ynth ~ ticks or ~ pong ~, o ~ l ~ ttr ~ 3 ~ 3 protluits n ~ t ~ real s ~ rvant de r ~ blanket t of pl ~ Sp ~ r e ~ rnple, de3 ~ els de coll ~ ne, cleR del ~ e4 ~. orl ~ lne ~ ni7 ~ 1a), L ~ compos ~ .tion FGP`-doxtr ~ n ~ ulfot ~ on ~ olutlon do ~ t puolr lmpr ~ n ~ r these dlf ~ n ~ orma ~ d ~ pon) 3 ~ nts, de monl ~ re que le fQ ~ t ~ Jr F ~ F pul ~ t ~ on contaet ou dlf f ~ lser s ~ ul or e ~ 3 o-; ie c ~ ud ~ xtt- ~ n ~ 6u fat0 ~ squ 'nu: c tls ~ u ~ clbleo. L ~ co7npo91tlon ~ lon l ~ n ~ ntlon ~ 3t lo ~ ppl ~ qu ~ e ~ lem ~ nt de m ~ ni ~ ro ~ moyol ~ tr ~ ~ slnt4n ~ 0. ~ ur le ~ ite de 1 ~ bl ~ s ~ ure et: sur le ~ b ~ er ~ oo ~ l ~ art ~ Y ~
m ~ lntlen de 1 hyclr ~ tatlon ~ lon lo ~ ~ ee ngq ~ os ~ e 1 ~ ho ~ m ~ du profession, p ~ rt ~ u ~ l ~ rem ~ nt ~ l ~ bo ~ d ~ n ~ le dom ~ ln ~ d ~ s graft ~ ~ c ~ pe, ~ u. Ie:; p ~ næ ~ ment ~ ~ ec u ~ lf ~ peuvont ~ tro lr / ~ pr ~ n ~ d ~ 1A m- ~ m m niere, ~ dsorb ~ or recover a ~ upport n ~ t ul ~ el ou: 3ynth ~ t lqu2.
Pou ~ le ~ ~ ppllc ~ tlon ~ sur 1 ~ 0 cvrn ~ o, 1 ~ v ~ h ~ cule dvlt ~ trR compo.tlbl ~ av ~ t.ol ~ ranc ~ cul41re ~ on c ~ t6 ~ r ~
co ~ nrne ~ mp ~ of such a v ~ lllcul ~ pro ult commc ~ -cl ~
:; OR 3 the name of ~ L ~ er l bu le ~ J), le ~ t on ~ lln ~
solut iVI- lsotonlq ~ eu: 3 ~ p ~ l ~ e:; ~ mpl ~ 40c ~ dron ~ 1 ~ 1erc ~
5h ~ rp-Dohm ~) or ~ equivalent), or oro ~ or ~ form ~
gel forming a ~ notch ~ t ~ ct ~ ur d ~ 1A c rn ~. C ~ 3 v- ~ h ~ culo.s p ~ uv ~ nt cont ~ n ~ r ~ 3alomen ~ dw ~ e ont ~ cn ~ orv ~ t * ur ~ comn ~, ~
3;: hlorur ~ s ~ ben ~ yldim ~ thyl 41kyl ~ mol lu ~ or f ~ thyl ~ nQ
di ~ mine t ~ tr ~ ac ~ t ~ t ~ 3 dl: sodium ~ DTA).
It can é ~ ra ~ nt ~ r61 ~ ouh ~ 1 tlble d 'lnclur ~, ion ~
de ~ llposom ~, une ~ s ~ oclclt ~ on Ft; F-dox ~ An ~ 4Ulf ~ t ~
hy ~ rosolubl ~ dc 1 'lnv ~ ntlon. C ~ 3 llpo ~, n ~ mauve ~ t dtr ~
qlorEs lncorpor ~ d ~ ns le ~ compo ~ ltlono ~: rita: ~ cl ~ e ~ u, 3, p ~ r ~ x ~ rnpl ~, cr ~ m ~: a, pc) mm ~ d ~ o, utll ~ 3 ~, ~ llo ~ qu ~ s or for the iMp ~; n ~ tion d ~, ~ upport ~, ~ omm ~ ~: ~ pan ~ m ~ nt ~.
n ~ la c ~ i3 o ~ l 'ut ~ lloo d ~ d ~ ctr ~ neo ~ ulf ~ te :;
lni ~ ol ~ ~; les, '~ eu ~ -cl p ~ uvont ~ itro s1mpl ~ ~ nt a ~ o ~ i3 a or bicr ~ talleo 4 ~ 0clatloao p ~ uvont ~ ro lnelusoi3 ~ sb ~
deo i3upporti3, ~ omme d ~ cr ~ m ~, de ~ ~ 4J ~ tln ~ o ou d ~, 3 gel: 3 de . ~:, '''''''"''''''.

,, '' :. . .. ~:, '' ;. ,,: ~

; Z'lJ ~; ~ q.'D2 (5 collr. ~ z ~ s, ~ 1 ~ be ~ ppltgu, '~, s fii ~ fl ~ r ~ ~ ynth ~ tlqu ~
ou rlatu ~ e ~ ~ f ~ ls cl ~ clure ?, or l ~ iF ~ n ~ r ~ upp- ~ rts habl.u ~ ls ~ - ~ ep ~ n ~ e. ~ ent ~ recover ~ m ~ n. Lnclu ~ lcn de ~ s of: ~ tranes sulfata3 ln. ~ lubles peut ~ iQ ~ lro p ~ rl ~ blbltlon ~ / or ~ dc ~ lti ~ nd ~ un ~: solutlon de coll ~ e ~ n ~ ~ t ~ llfl ~ ~ tLon.
On p;: ut notamm ~ nt ~! Ti ~ l ~ er le ~ ot ~ 3 op ~ e ~ t31re d ~ crlt 4 ~ n ~; the tre- ~ t U5-A-4 0 ~ 0 0. ~; u ~ v4nt lsquel nr ~ llse ~ In ~
co ~ nposltion ~, t ~ lflé * ~ omp ~ slt ~ d ~ ns ~ u ~ llo l ~ p ~ rtl ~ an CC`t ') tACt nvec la ble ~ ure a ~ t ~ ~ coll ~ n ~ r ~ tlcul ~
~ 0 ~ \ Vf ~ n ~ lyco ~ mln ~ gly ~ n ~ SI ~ n ~ 4tant o ~ teflu par , ~ d 1.11 t 1 on d ud 1 tgl yc oo ~ ~ 1 no ~ 1 ye anh ~ uc 1 1 Qs ~ n ~ ~ Oa ub ~ nslhmbl ~ ~ both pr ~ Iplt ~ or r ~ t1cul ~ p ~ t ~ du ~ lut ~. ~ rolddhydt ~. '' By 0111 ~, co ~ nme ~ ndlqu ~ ~ -dos ~ ll3 ~ le: ~
;! ~ C ~! RlpO ~ n ~: 3 ~ 1 ~ n 1 'In ~ nt1on p ~ uv ~ nt ~; 3ol ~ 9e ~ t p ~,: s. ~ nt,: l ~ 4u ~ un ~ f ~ r ~ e appl $ cable lc ~ letn ~ nt p ~ r ~ ol ~
Interr *, for example c ~. ~ M ~ pl ~ tr ~, n ~ 4tro-ont ~ rlqu ~ 6 cu pO ~ b ~; lyophll15 ~ .ea ~ nclusas ~ u nan d no das z ~ lule3.
LA pr ~ pdr ~ tlon de ~ co ~ po ~ ltl ns de tr ~ lt ~ merlt, ~
ic, ~ trl. ~ 4t ~ 0n: 3e10n Ll LnV ~ rlt10n ~: 3t, d Un ~ ~ n ~ g ~ n ~ r41 ~, r ~ e ~ elon de3 m ~ thodQra cla: 3 ~.; lqu ~ blan co ~ nu ~ 3 ~ n nl4u ~ e. L ~: ~ dos ~ $ ~ ~ d ~ nlnl ~ tre ~ ol ~ ur fr ~ ql ~ ence .sont or1 ~ 3 not ~ mtner ~ t: se10n 11ImpDrt ~ t. ~ d ~ l ~ pl ~ lC.
Enflrll ~ COi ~ tpOi31t10lll ~ 5 ~ 310 ~, 'ln ~ / entLon kl ~ or. ~ Ent lert ~ ent, 3pplic ~ t lot ~ t dah ~ l ~ dom ~ lno o ~ m ~ tl ~ ue, for ~ m ~ IIor ~ r ~ t4t d ~ l ~ p ~ au. Ln ~ ttnt on ~ nc ~ o ~ m ~ n ~ s ~ mpo3 ~ tlon ~ co ~ t ~ tn ~ nt, n ~ un ~ uppt ~ t cosm- ~ tlq ~ emerlt ~ c ~ lpt ~, une a6 ~ io ~; 14 lon pr ~ clt ~ FCF--tran ~ ~ ultat ~ :, c ~ tte c ~ poGlt ~ on s ~ pr ~ s ~ rltAnt not ~ mrnent, i ~; form ~ key ~ tlor, ~ al or cr ~ t. .. ....
We ~ crlr ~ molnt ~ nortt l ~; ~ of ~ t ~ d ~ d ~ tl- ~ tn ~
6ulr ~ t ~ UI ~ prot ~ ct ~ ot ~ 3 1 '~ itl ~ l ~ t bLolo ~ l ~ ue d ~ 3 rt ~;.
L ~ h, ~ v ~ lu ~ n ~ 1 Y ~ t ~ ctl - ~ t - ~
h ~ ue l ~ 'CF3 s ~ 2nt d4c ~ 1te, 3 d4n ~ dc no ~ nbreuo ~.
~ IL> lic. ~ Tlor ~ t ~ lle.3 ~ have everything ~ ba- ~ olt ~ r un ', ,,', '",'. i '' .1,,, ' Z ~

l ~
m ~ .lsuro (.! el 'b ~ lgrrlent4t l4n, lu n ~ br ~ da C l ~ u1 ~ 3 ~ nd ~ p4 rl ~ :, d- ~ ..e :, crol3is4nte ~ C1 ~ 3 farteur6 4 ~ 10ut 4 ~ u ~ place of cultl ~ r ~ d ~ llule ~, ~ olt ~ ur un ~ 'nt ~ t ~ on do 1 'lnl: orpo ~ tlon de thymldine t ~ ltl ~ ed ~ ~ DIl de ~ cellul ~ J
3t Irnul ~ r: ~ 3 p ~ r lr ~ f ~ cte ~ rd ~ crol6 $ ~ ne ~. D ~ ns 1 ~ u: c protocolc, these ~ u ~ ent ~ tl ~ n ~ on ~ d ~ p ~, d ~ nt ~ de 1 ~ d ~ d ~
f ~ ct ~ ur ~ jo-lté ~, ~ t 1 'we can ~ unc et4t Lr de ~ ef ~ ots d ~ e ~ t cl ~. ~ cour ~ e, ~ do ~ r ~ pon ~ e ~ voc un 4f ~ It ~ xl ~ l dQ
~ klrn ~ l ~ tl ~ n. P ~ r ~ irnpllflc ~ tlon, on d ~ lnlt IJr, e unlt ~ de 1 ~ st in; ll ~ t lon conune 14 do ~ 3e d ~ f ~ ct2 ~ rd cro ~ 3 ~ nc ~ qui, ~, ~ out ~ n ~ r ~ d ~ u do e ~ t ur ~ ~ ur d ~ r ~
cell ~ cLble ~, ~ st cop ~ blo d 'indulrs n ~ nug ~ n ~ cSlor, du no ~, br ~ df ~ c ~ llule ~ c ~ ud ~ lnco ~ por ~ tlt I d ~ 3 thymldln0 trlt11 ~ corre6por ~ nt ~ molt1 ~ 0%) d ~ ln v ~ l ~ ur r; m ~ 3 ~ lm ~ 3 dl3 Cl5! ~ u ~ t. ~ nt ~ t lon m ~ jur ~ ~ na; 1 ~ ~ ourb ~ d, ~ s, ~
r ~ pOI- ~; 3 ~ C ~ 3ttQ, i;! ~ flnl ~ lon ~ t lo r ~ pr ~ cl ~ tlblllt ~ d ~ c ~: ~
m ~ sur ~ s .ont expo ~ 3 notan ~ m ~ nt p ~ Plc l ~ t ~ t ~ oll., ~ n3 ~ C ~ lIul ~ r ~ nd Mole ~ r Blology,; ~, p ~, ~ 5-I I0 (1994 ~)).
A1A ~ 1 ~ plu3 1 ~ unl ~ 3t m ~ ntenue ~ un v41 ~ ur ~ lbl ~, plu3 f ~ c ~ eur con ~ r ~ une ~ c ~ iYlt ~ blc> lo ~ q ~ e lmport ~ nt, 3.

A - ~ f ~ kr ~ ~ d ~ u ctl ~. d, 1 ~: 3 EF. ~ .d ~ t ~! ~
S ~ ~ and P ~, ns.
. . .j l ~ n: ~ c ~: ~ xp ~ rlence ~, we use 1 ~ 8 of ~ Golutlonr ~ -m, `-res da F: GFo, c4ntennnt 1 ~ FGF ~ ~ n: ~ ILlt ~ on ~ un ~
onr r ~ t ~ ~ r ~ tl ~ n ~ ~ S ~ p ~ r rnl llllltr ~, ~ lno un t ~ mp, ~ n r! h ~ p ~ l ~ ta lsotoniqu ~ t ~ B ~ t ~ ont ~ n ~ e ~ up ~ s ~ ud ~ xtr ~ no . "l ~ l î dL ~ tDS) 1 ~. 63 ~; 3 ~ Ju cdtdlogu ~ SI ~ ~, Jl 400 J ~ ml. 1.
dc1 ~ u ~ a ~ lqu ~ utlll ~ vr ~ t ~ 1'Q ~ n ~ bovin6 et tr ~ lts du c ~ r ~ de bo ~ ut, aclorl 1 ~ p ~ lblIc ~ t ~ on d ~ Court ~
~ u u l. IiQnJ ~ BlvchlM1 ~, 69, p ~ $ ~ '~ $ II ~ 87) ~.
10, ul ~ 1 ~ c ~ tt C ~ iol u ~ lon are pr ~ v ~ s ~ t nl ~ l4n ~
.3 ~ it ~ 1 ml ~ te- ~ pon ~ EiS ~ pH 7), ~ lt ~ C ~
; J ~ kti ~ ll * ~ 4u ¢ n ~ Ju ~ t ~ 4 pH ~ ~ nrlron iN ~, ~ olt b I ~
: ~ o ~ lt ~ ~ N; ~ O ~ 11ul ~ *, ~ 3u.st ~ e ~ pll ~), G '. C ~ s ~ ch4ntll1t) l ~ s (~ nt . j ;. ';'','' ~ '''';

~ "'''''~,,',~; li, ~.,:; ~;,;, ~ '~," ,, ", ~"""

Z1, ~, ~,? ~ `.54 lncub ~ e ù 2 ~ 'C p ~ nd ~ nt 2 h ~ ure ~ and a 1: ~ v ~ m ~ nt d ~
e ~ t ~ t'f ~ l-tu ~ for the back ~ ge of the actl ~ ~ blolo ~ l ~ u ~.
Fl ~ sour 1 pr ~ sel ~ te 1 ~ co ~ b ~ os ~ -r ~ pon ~ tu ~ FI, ~ s ~ rd ~ flbrobl ~ te.Y d ~ ~ ertn ~ hun7 ~ n .tA ¢ 1523) nde de 1 ~ f ~ r ~ I
Ab6cl ~ 3 ~ es: do ~ es ~ ef ~ ct ~ ur de cr l ~ s ~ n ~ e (p ~ / ml ~
~ r ~ onn ~ polJrcent4 ~ ed ~ 3tl ~ ulo lon 1 ~: bFCP ~ DS ~ pH 7 ~: bFGF ~ p ~ 7 V: bF 5F ~ ~ S ~ p ~ ~
A ~. ; b ~ OF ~ pH 9 o: bFGF I ~ 5 ~ p ~ l 2 : bFGr: ~ pl 1 2.
~ 5 L'~ ugment ~ t ~ on ~ 'incorpor ~ tlo d ~ thym ~ tlr ~ e tr ~ t ~ r ~ pre ~ ent ~ 1 ~ v ~ their number 9 eoupo p ~ r mlnute ~ cprn ~ obt ~ nude au plate ~ u de 1 ~ ceurbo d * ~ -r ~ pon ~ e ~ u ~ I: G ~ ' selll m ~ lns la v ~ r en epln ds thymld ~ trlt ~ lncorporee don6 le, 3 cell ~ en ~ b: ~ -an :: e ~ e FG ~ et ~ rmlné ~ d ~ n3 l ~
~) same ~ eXp ~ riqnee ~
Le T ~ ble ~ ulr ~ 3ume 1 ~ $ r ~ Ssull ~ t8 obt ~ nlJ ~ ~ ve ~
al: ~ t '~ t bt'GF, L' unlt ~ i de ~ tlmulation ~ t ai ~ bltralr ~ ment dorIn ~ ~ L for 1 ~ 4 ~ GI: or the bF ~ F ~ oc ~ dp ~ rt ln ~ ub ~ hang 2 ~ ur ~ 3 ~ Z ~. . . .

Tqk ~
_____ ~ _____._____ ~ ___ ____________ ~ _ ~
~ _____.___.___ ___ bl; '~ F ~ O'C). 9 ~ 1: 0 ~ (2 ~ 0- ~) S: ~ 13 3 ~ b ~ qF ~ DS ~ 2 Ii, 20 ~) 1 ~, 5 ~ FCr ~
o ~ 'OI;' ~) 'C) ~ G
cl'OI ~ + C ~ S ~ 2 I ~, 2 ~) 'C) ~, 5, ~ 2 ~ _ ~
: ~ 5 ',. , I, -i., ,. ,. ..: ~, ''. ',. :,, ~. : '.' , '! . : ''. .
'. ,:; ',,. '',, '''''I,'',,!':

', Z7) ~ ' {~ - Etf. ~ T! ~; T ~ n / ~ f ~ te !! ~ ~
p. , ~ ure ~ .t ~ i ~ ~ ~ h ~, C ~ ' Dat) 3 c ~ tt ~ p ~ rlenc6 ~, le ~ 0 pr ~ p ~ re Comm ~ ~ u poi.nt A ci ~ ~ e ~ slls, for the form ~ cid ~, at pÆr ~ recctmb ~; on ~ r ~ tlque pour la form ~ ba ~ iquçl d 'orl ~ In ~ hu ~ ln ~, ~ chet ~
~ upr ~ de 1 ~ 5 ~ clét ~ A ~ n, ~ rs ~ hsn ~ ~ u Cllnl ~ c ~ nc ~ ~ P, ~ rl ~), C'S4 Lncub ~ b 4'C, ~) C, 37'C or ~ O'C p ~ ~ d ~ nt d ~ erent ~ s p ~ rloul ~ s, in 1 'Qb ~ C ~ or in pr ~: ~ enc0 c ~ ~, 00, l ~ z; ~ ed ~ xtr. ~ ns u ~. f ~ t ~, p ~ d ~ D ~.

~ L ~
_ ~ _ ~ __________-_____, _______________ ~

4'C 0- 37'CGO'C
___________.____________________ _ .__. ~
. 'j bFCF t ~ 4. ~ nn 1 I ~ F ~ iF t = 3 ~ r.nn 1 1 3, 5 ~; :) O
b ~ 'Of: ~ DS t - 30 Ir.n 1! I
aFGF t Y ~ ml ~ l , ~ FCl ': k ~ 30 n ~ n II ~ ~ l os ~
, 'O ~ F ~ F 1- [~ S t = 30 rR ~ O. 4 O, ~ O, 4 bFGF t - ~ 4 h ~ Ilre: 3 l I
bl; OF t DS; ~ ~ 4 hrs @ ures I 1 ~ CI: I 1 ~:; F ~, 4 t :), 4 O, ~ S bFGF t ~ 7 ~ bear Z! 3 ~ lOO
bFCF ~ D ~ t = 7 D our ~ I 1 aFGF t ~ 7, ~ YES ~, 5 ~ ~ lOO
~ r: ~ F ~ ~ st - 7 ~ our ~ 0, 4 O, 4 3 . _ __ ~ ______________ Lo ~ donn ~ r ~ pport ~ $ d ~ n ~ e ~ tobl ~ oux I ~ t ~
lllu.ctr ~ t1t l ~ ff ~ t protectBur tr ~ procur ~ p ~ r le ~ sùr the F ~ t :. Aln ~; l, toutQ l ~ octlYSt6 blolo Iqu ~ du bFGF ~ t corl ~ e ~ rv ~ ctpr ~ $ 7 Days of ln ~ uba ~ tlon ~ 37'C, ~ l ~ rs ~ que ~ .n 1 '~ b; s ~ ncr: de t) S, none ~ ct ~ t ~ n ~ ~ t ~ iL d ~ t ~ ctobl ~.
3 ~ ', , :. . . ::
. ... .
?
... ...
~,. , E ~ EI ~ PLI ~ r ~ 'APPL ~ TION IN VIVO:
p ~ l- l men to l. ;
The op ~ ro ~ lons reoll ~ e ~ ~ ur dt ~: sr ~ ts W ~:, t ~ r m ~ le ~ 3 de 300 t ~ 400 ~ r ~ mme. ~. Ch ~ qu ~ exp ri ~ nco t3 ~ f fect ~ e sllr ~ Jne st ~ rl ~ d ~ 5 ~ nim ~ u ~.

~ ep ~ s;
We prstlq ~ e two typo ~ dt ~ pll- e ~ cut ~ and ~ ~ ur 1 It: ~ ft ~ ce d ~ rs ~ le, p ~ ol ~ blem ~ nt r4s ~ e, t ~ r ~ lmt ~ u ~.
1) Ie: ~; pr ~ l ~ enements ~ t ~ nt ef ~ ectu ~ 3 p ~ a emportQ-p ~ 4ce o-~ punch (0.6 crnl di ~ m4tl ~> 3usqu ~ plsncht r rnu ~ culalr ~.
2 ~ DQ3 ~ C srlflc4tion. ~ D ~ 1 cm long 00nt prt ~ tlqu ~ e ~ eu calpel. It. ~ N '~ f fc ~ cte ~ t que ~ 1 ~ rt ~ Slon dor ~ o-Spl ~ t rmiqu ~. .
.j. . .
Summer ~ and:
S ~ lon 1 ~ typ ~ d ~ pl41 ~ s, le3 le ~ ur ~ 3 .s ~ nt tr ~ ltee ~ p ~ r dlff ~ rent ~ m ~ lan ~ dl3 pro ~ lts en ~ olutioll ~ l ~ ns du: 3lrum phy ~ iolo ~ ue t ~ lnponn ~ ~ 5 p ~ 71 4 ~ ~ t ~ rllls ~.
Ce ~ 301utioll ~ 30nt d ~ p- ~ k ~ for 1 ~ 9 pl ~ ap ~ r punch d ~ n ~ a t ~ mpon d ~ eoll ~ sèn ~ n ~; ~ tc ~ t ~ pr- ~ d ~ bcoup ~ ~ u: ~.
"j rn ~ urc ~ e: c ~ etes de 1 'xcl ~ ion ~ l ~ 3ul ~ 1r.
D ~ n6 the ca ~ of ~ ~ c ~ rlflcation, ~ 8 pl-oduit ~ ~ have d ~ pos ~ l3 dlrrCte! mqnt; i3CUei form @ llqulda ur la pl ~
I..e ~ effcto de ~ 9ocl ~ tlc; \ n F ~ <b ~ lqua ou ~ clcl ~
au un m ~ l ~ n ~ ~ 3n ~ olutlon d ~ I n ~ ~ ~ 0 ~ ml ~ ~ t ~
d-3xtrane4 ~ ulf ~ t6 ~ n ~ olutlon sl ~ lO0 ~ ~ I m ~ / nil ~ 30nt ~ v ~ lu ~ and eomparlSo ~ actlon te ch ~ cu I ds ~ s ~ on ~ tltu ~ n ~ s, ct) nsld ~ r ~ 3 ccmn ~ Témoirl3 d ~ r ~ ction ~ C llh ~ Olutlon de d ~ o: 301ul: ~ 0 ~, FGF, dextr4ne r ~ ulfut-). L DS l ~ te ~ lt ~ QlU
~ i ~ ponlbl ~ chc ~ S ~ ÇMA ~ 0 ~ 5 1 ~ r ~ f ~ r ~ nc e ~ s ~ 3.
.i? s ., '.'.

, ..,.

., '', ','. , ''. '''''.

~ l Z ~ r5 ~
, "

~ h ~ que s ~ rl ~ experiment ~ le d 's lm ~ ux ~ 3t ~ 1 ~ c ~ lf L ~ -3 b lnt ~ rv ~ lles d ~ t ~ mp ~ d4flnls p ~ rp ~ rlo ~ de ~ 4 i-eurQs, ~ t 1 ~ re ~ ions 1 ~ 6Se3 ~ have p ~ high ~ e ~ for cl I ~ X t ~ p ~ 3 of ~ studies:
1 ~ a ~ ~ n ~ ly ~ e morphologlq ~ le ext ~ -n ~ ~ ~ e pl4r ~ ri ~ de 1 ~ pl ~
2) ~ no 6 study hlcto ~ oglque.

Result ~ t ~;
. ~ 1 ~ ~, vlyo Ç ~. ~ ~ ~
bu F ~ F m ~ rqu ~ rnt ~ o ~ ctlven ~ en ~ l 'ls ~ de I 125 ~ st Dépo.s ~ en p ~ nce ou ~ n ~ '4b ~ ence d ~ Il; tr ~ na ~ aulf ~ t ~ a cl ~ ns ~ nt ~ mpen d ~ coll ~ ne ~ according to l ~ p ~ otoeol ~ d ~ crl t by l ~ o ~ nner 1 ~ 1. et al, d ~ ns ~ Proc. Natl. J ~ C ~ d. Sc (US ~
pOZ ~ -5 'jo ~ -502s ~ ~ 9B ~>) ~.
L4 v ~ ri ~ tlon de l ~ rodlooctlvl: 1 ~ d ~ n ~ a le coll ~ g ~ n ~
`~ irnprlSy; n ~ e ~ t ~ ppr ~ cl ~ e ~ n funct ~ on tu te 3np ~ <f ~ su ~ rs 2).
717 ~ L ~ d ~ 1 ~ f ~, J ~ e Abc ls ~ t emp: 3 in h ~ 3ur ~
~ rdonn ~ eY ~ 18 ra ~ oe ~ ct 1 ~ t ~ m ~ uré ~ p ~
report ~ l ~ r ~ dio ~ ctlvlt ~ el 'C ~ lgln ~
bFOF ~ DS
., ~ 5 coll ~ g ~ na x --- x bFO ~
bF ~ 3F
2 cm ~ ol ~ l plol ~ {
bFOI ~ ~ ~ S
Ln r ~ dlt ~ ctlvlt ~ ~ st me ~ ur ~ ~ n ~ 1 cla collt ~ n ~ e ~ d ~ n6 de ~ pr ~ l ~ v ~ DleAtl3 ~ ff ~ t ~ t41 ~ ~ np ~ ph ~ rle do l ~ pl ~ c, h ~ crn de ~ cllc-el, p ~ r un pun ~ h Ibquivol ~ nt celu ~ d 'c ~ rl ~ slne.

; ~

2l ~ 0 ~ 4 II - E ~ ~ orp. ~ Qls ~ IF ues ~ ~ 1 ~ 3n ~ L5U ~.

A) ~ goose ~. phr emp. ~ r ~ pi ~ ,; .
~) Morphological study t L ~ ob ~ ervl ~ tlor d ~ volut ~ on ~ ~ pl ~ o ~ ~ 1 'eye nu perm ~ t de c ~ n ~ t ~ t ~ r an actlor t ~ r att ~ ~
1 '~ s ~ o ~ l ~ tion FG ~ ~ cloxtr ~ n ~ ~ ulfQt ~ ~ u 14 vlt ~ and the qu ~ l ~ t ~ de 1 ~ c ~ c ~ trls ~ tlon super ~ lc ~ el ~ ~ pldcro ~ tl ~ n lys ~ du ~ lllot).
1 ~ 3, 1) After 24 hours, the ~ tAmpono of co3 l ~ n ~ ~ rnpr ~ n ~
p ~ re ~ tte ~ socl ~ tlon ont tot ~ lamær ~ ~ dhkr ~ aux p ~ rols d ~ 1 ~ pl ~ ie, and can ~ tr ~ pr 1QV ~ only p ~ r 1 ~ 310n tls ~ u ~ r ~ r ~ s. Le3 exp ~ r ~ e ~ t ~ tlon3 t ~ oin ~ n ~
m ~ nlfe ~ t ~ rlt uno ~ h ~ r2nc ~ tot ~ le d ~ co ~ n ~ que ~ u after ~ 3 ~ ~ 48 hours ~ ur ~.
2) The ~ r ~ plthéllo ~ tlon ~: ~ t vl ~ l'ole ~ 'o ~ ll nu ~ u bout a 3 ~ ml3 day before the ~ ssocl ~ lon FC ~ ~ of:; tr ~ ne : iulfate, ~ lo ~ - ~ q ~ 'ur ~ e Im ~ a ~ ei ~ enti ~ le Igu ~ le ~ m, ~ ln ~
request for: 3 t ~ mp ~ of experience ~ tlo ~ 7 ~. ~ ur ~.
3) / ~ n ~ ly ~ pl ~ nl ~ n ~ trlg ~ In4 ~ lySl ~ tl ~ nim ~ trlqu ~ ao 1 ~
~ urf ~ cc extel-n ~ de ~ pl ~ moil ~ rQ 'Qbsenc ~ t ~, gole de r ~ tr ~ ction ~ o ~ t ~ su: ~ ~ nr ~ n ~ r ~ l: on.
2 ~ L ~ t ~ bl ~ aux 3 et 4 r ~ pp ~ r ~ on l ~ o moyen ~ e ~ de five e ~ perlencasl ln ~ thrilling ~ conc ~ rn ~ t:
~: Le r ~ appor ~ ~ xprlbnb ~ n pou c ~ rlt ~ ge ~, d ~ la 3urfacc de 1 ~ clc ~ trlco t ~ l ~ ln ~ de L ~ - surf ~ ce de la cic ~ trlec tru ~ t ~ p ~ a dlff ~ rent ~
3v o ~ $ ~ ci ~ t lons.
.Q ~ ,: Le roppol-t du ptlrLmbtr ~ t ~ ~ h ~ gu ~ cic4trlc ~
~ ec la ~ urf ace de c ~ 3 m ~ m ~ ~ ci ~ trlca ~.
., ~! j '~.
. ' 2i ~ .54 1 ~
L ~ s ~ ocl ~ tlon FCF ~ dextr ~ n ~ 8 lf ~ te d ~ n. ~ Le 3 ~ pport coll ~ ne ~ T ~ ble ~ u 3) permee0 clc ~ trl ~ tlon plus h ~ rmo ~ leuse, pr ~, 3entnnt a fo ~ ble r ~ tr ~ tl.on ~ es t ~ 6suo ~ 102, 58 '~, ~ u Day B, center ~ 49, 27% ~ V ~ I: 0 ~ ~ -Yl).
~ ï De plu ~ 'a ~ p ~ act clrc ~ lolr ~ ~ la pl ~ le ~ e eons ~ rv ~ d '~ ut ~ nt ml ~ ux, ~ u COUI ~ de l ~ c ~ trl ~ otl4n que le r ~ pport p ~ rlr ~ tre P t ~ 1 ~ pl ~ le ~ ~ urf4c ~ d ~ plDle exprl ~ dQn ~ læ T4b ~ e ~ u 4 ~ ot plu ~ f ~ Lbl. LA Y ~ r dl3 C ~
r ~ pport perm ~ t cQr4ct ~ r ~ ser lel confor ~ tlon ~ e 1 / ~ pl ~ le 1 ~ in cour3 ~ a cic ~ trl3l ~ t lon. L ~: 3 vQl6 ~ piss o ~ more ~ f ~ ble ~ s are enre ~ l st r ~: 3 p ~ r 1 '4 ~ socL ~ t lon F ~ P d ~ xt r ~ ul ~ te dan ~ l ¢ q E; 81 ~ de ooll ~ z ~ ne. ~ in31, ~ 1 ~ Jur 8, the rCIpport P / S ~ td ~ 0, 22, ~ lor ~ que, pou ~ le FG ~ ~ ul, la v ~ eu ~ e. ~ T
0, 3! i ~, ~ en ~ l ~ lem ~ nt l ~ enti ~ ue ~ c ~ llo bt ~ r ~ u ~ dor ~ - 193 l ~ Témolns sans colla ~ s ~ n ~. The ple ~ t ~ m ~, h ~ clc ~ trl ~ en ~ p ~ r r ~ pprochement de ~ re ~ and prennoAt w ~ p ~ ct ov ~ î elm.
Ce ~ l est ~ 3; ule ~ ent lllu ~ l: r ~ d ~ n ~ 1 ~ 3 phC: o ~ r ~ phle ~ de ~ f ~ ce4 dora ~ l ~ 3 ~ nln) ~ u ~ tr ~ lt ~

~ O ~ q ~ B le t ~
5urf ~ c ~ d ~ pl41e t ~ n ~ oin (- ~ 3olutlon F lyololoP: lqlle) ~ ___ ___ .__ _ ____ Product ~ ~ oll ~; ~ nq Col ~ ~ 3 ~ n ~! Collu ~ ne Col laé; ~ sne + I ~ S ~ ~ F ~ F ~ b ~ F ~ ~ S
~ ul 50 ~ g ~ ml I ~ sOm ~ t Days 500 ~ 6 ~ ml ~ - -2 80, 7 ~ 1; 1, 43 7, C ~ 0 3 ~ 0, 43 9 '~, 54 1 ~, 127 174, 42 g ~, 79 77, 08 ~ 5, II 1 l 7, 24 3V 8 92, 409 ~, 14 ~, a7: lO ~, 51g ___ ________ ~ ________ _ _ .__ ~ __ _ ~ ~ _ _ _____ _ __ 3g '..' - ~: ~

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R ~ ppGrt F ~ / ~ du p ~ tr ~ ~ e 1 ~ pl ~ le: ~ u 14 ~ urf ~ c ~ ~ c ~, t ~
pl al ~.
~ rodult ~ Gc ~ g ~ n ~ Coll ~ no c ~ bn ~
T ~ oln Col ~ e ~ b ~ O ~ ~ ~ S ~ FOF
~ ns I t ~ i 000 ~ 5 5C10, ~ lg ~ t ~ v ~ r ~; coll ~ nQ ~ I ~ g. -, ~ ~ __ ____ _ _ _ _ O, 1 ~ O, Z2V, ~ 5 C ~, 20 0, 19 2 ~ 0, ~ 10, ~ 5 0 ~ Z ~ C ~, t ~
. ~ 0, 22 0, 360, 16 0, ~ '6 0, 1 4 0, 3 ~ 0, 20t ~, t7 ~ Cl, t ~
6 0,: ~ 7 ~ 8 0, ~ 5 C ~, 15 ~, 7 ~ ~, 4 ~ 0.39 C ~, 4 I .5 b L ~ sr ~ lonc ~ tr- ~ lt ~ e: 3 ~ 30nt pr l ~ v ~ o, flY ~ r ~
lMr ~ rt! 3n, $ ~ np ~ ffl rl ~. L ~ t ud ~ hio olo ~ st r ~
'- ~ u; ~ Ollpii ~ ia d- ~ 7 Jlm. The; color ~ tlo s utL11 ~ 3 p ~ rM ~ tt ~ nt d ~ tud ~ 3.s topo ~ r ~ ph1qu ~ hl ~ tochL ~ lque ~.
the ~ fig ~ s ~, 4 7 are d ~ p oto ~ r ~ phle ~ ~ ro ~.
d ~ up ~ h. ~ tolo ~ qu ~ 3, obtained ~ ~ 4 o ~ r ~ i for 1 ~
î i, ~ 5ur ~ 3, 4 5, Jour ~ pour 1 ~ flsur ~ 4 ~ ~ 3 Jour ~ pour le ,, '5 fi ~, u ~ 5 ~ 7-Flg. 3 ~ 0) r ~ prb ~ nt ~ r oult ~ t ~ ~ bt ~ nu4 ~ rc u c411 ~ n ~ ul ~ t ~ ol n ~ ~ t ~ Pl ~. 4 t .; 25 ~) r ~ pr ~ r ~ t ~ le ~ r ~ U1 ~ 4t ~ $ 1m ~ 1 ~ 1st with 1 ~ ocl ~ t ~ or ~ ~) S t PGP ~ an 1 ~ ~ o ~ n ~ t ~ - ~ F ~ S. 5 SX S003 repr ~ nt ~ un d ~ l 411 d ~ o ~ u; a ~ n r ~ n, ~ r ~ t lon col-r ~ l ~ pord ~ nt 3 14 ~ l ~ ur ~ t 4.
Fl ~ $ ~ 6 and 7 ~ 7 ~ 2gO) reprbsentan cl- ~ r ~. ~ Ul ~ 4t ~
~: lc, ~ t ~ tlon. ~ d ~ pl ~ le: ~ obt ~ n ~ J re, ~ p ~: tlv ~ m ~ nt ovec a ~ ~ Otl.Jt lon phy; ilûlozlqu ~ t ~ on ~ t ~ v ~ a oc l ~ tlc, n ~ FCiF ~ tr ~ n ~ sul f ~ ~.
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~ CV aY'X ~ rox T ~ l ~ copl ~ r 702U, 10- 4-89; 11:49; 33 1 874 51 37 ~, ~, S &H; ~ 1 . . .
"'2q ~ .. 3 ¢' ~ i5 CO
V ~ n3 1 ~, ~ xp ~ rler ~ cec ropport ~ ~ nr ~ f ~ r ~ anc ~ au ;;
s 3 ~ S, we have utllls ~ ~ es m ~ e ~ o ~ pos ~ tlons qu ~
c ~ lle ~ lndl ~ u ~ ut ~ ble ~ u 4.
L ~ an ~ ly, ~ h ~ stolc, gique d4D ~ on ~ r ~ q ~ e 1 '~ sl ~ ocLatlon ~ + VS accelerates the ét ~ p ~! A tr ~ dlt10n ~ lleQ of 1 elcatrlsotion d, ~ rrn ~ - ~ plderD ~ eu and eua: Dnte 1 ~ qu ~ de.
t ~: 3us reconstl ~ u ~ - ~ p ~ r ratio ~ xo ~ ~ nt ~ tlons t, ~ oin.s (~ oll ~ 3 ~ ne sf ~ ul ~. A ~ nsl, in pl S ~ -lnc6 of F ~ F +
~ e: ~ trane, a ~ colonl ~ t lon very ~ rapld ~ ~ I Day ~ from ~ a c ~ t ~ gorleO c ~ lalre ~ ~ n ~ lronnante $ ~ 1l ~ obl4s ~ eo, sru ~ cul ~ LrC3 lls ~;) e:, ~ o ~ s ~ rv ~ e ~ por Sr d ~ st ~ ssUs ~ ln3 ;. ~ Lns, et, ~ n partlcull ~ r, ~ p ~ rt ~ r du con, ~ onet1 ~ ~ u pl anc her mus ,: ulo 1 re ~ str ~ 3 ~ c ~ nt.
~ an ~ m ~ k ~ mps ~ fl ~ ure 5, a n ~ o4n ~ iea ~ en ~ sQ
1 ~ allows ~ u tLss ~ - in f ~ r. ~ Atlen to be co on ~ p ~ r a ~ .nslt ~
t ~ for., ~ de capill ~ ire ;; ~ an ~ sulns ~ d ~ l ~ n ~ p ~ r le ~ hlffre ct ~ r ~ fbrence 10 ~ ur cett ~ fl ~ re), d ~ ll 1 ~; t ~ s.su: ~ en r. ~ n ~ r ~ Slon dk ~ ign ~ s by le ehlffr ~ ldr ~ fkrenc ~ 11. / ~ u L ~ ou ~ dc ~ ~ our: ~ ~ contr ~ 5 h ~ for the ~ t ~ oin ~
.0 r ~ plth ~ l ~ 6 ~ t ~ on cngagke ~ partlr t ~ 1 ~ pldeeme d ~ re ~
J ~ 1 ~ pl ~ lc arrives ~ confronts ~ ent. The father is S,;, ~ t, 31em ~ nt réor ~ an ~ s ~ ~ 3t il pr ~ entQ u ~ d ~ nslt ~ norm ~ l ~
~ f.3u ~ -e ~, 4 ~ 1ne J ~ r ~ cor ~ .p ~ otlveo ~ ent ~, IX tg3toln ~ t 4 1 d ~ n ~ lt ~ c ~ llul ~ b, ~ uco ~ p plu ~ f ~ l ~ re 3 ~, 5 5 ~ .ne ~ our ~. Ce ~ meme3 l ~ lu ~ tr ~ tlor ~ ts r ~ nt 1 '~ b ~ nce d ~
r ~ tr ~ t ~ on d ~ s bor ~ s dQ 1 ~ plzL ~ powr ~ t8 punchs ~ t- ~ lt ~: 7 p ~ r assocl4tlon F ~ F ~ D5, contr ~ lrem ~ ne ux t ~ olns qul C ~ th ~ 3nt pat- r ~ tr ~ ctlon le. ~ Tls ~ u3 exc 8 ~.
The clc ~ tr ~ sation of ~ plala ~ b ~ onue ~ p ~ r scarlflcatlon ~ t ~ valu ~ ep ~ ru ~ e ~ tud hlotclo ~ lqu ~
.3ffe ~ tu ~ 5 Jour3 ~ pr ~ s ~ 'op0ratlon. ct traltQmer, tt con ~ l3te 2n a ~ ~ eule ~ ppllc ~ t ion top ~ eu ~ tJ nl ~ o ~ td ~ 1 ~ le ~ lon l ~ a - m ~ m ~ of the opkt ~~ tlon. L ~ sr ~ sult ~ .o ~ have ~ r ~ ~ u ~
1 ~ 5 tl3ur ~. ~ 6 and 7 c ~ u $ mont ~ nt le: a tl ~ iiu ~ de 1 ~ p ~ y ~ nt ; ~ 5 received a ~ s- ~ lutlon phyJ.lolo ~ lqu ~ t ~ mo ~ ~ r ~ t 1 ~ aJo ~ t Ivn d ~ b ~ GF re,: om ~ 1n ~ nt ~ de ~; r ~ n ~ ~ ulf ~ e <~ n, ~

,.

R ~ V ~ r: x ~ rox -lel ~ copler ~ lu ~ u ilU ~ 51; 33 1 ~ 74 51 37 ~ q, S & H: #

Xl .r: ri ~ ~ t 5 ~ e cle: ~ tr ~ n ~, ulf ~ te pD ~) for 1 r L ~ ru ~ ~ l '.' .
'1 e ~ t ~ n- ~ t er. ~ Ue the presence of 1 a ~ scc S ~ tl Gn lndui ~ 1 'er.clench ~ ment of: a mkcarilsme3 c cntrlciel ~ qui ~ etnblent totnlem ~ nt t ~ rminé3 au 3 ~ n ~ e Jo r ~ uiv ~ nt 14 bl ~: 3on ~ ure ~). L. ' ~ plder ~ e p4r ~ P: i ~ aent recoR ~ t ltu ~
~, urn ~ have a ~ erme ~ ns ~, prS ~ nt ~ n ~ W ~ l ~ orgMnl ~ t lon 1r ~ dl:, cerna ~ 1e des tlssu3 envlronnDnt. ~ nr ~ v ~ nch ~
cl- ~ atri ~ tlor ~ ~ u té ~ l ~ oln Cflgur ~ 6 ~ t ~ r3 las o ~ st ~ 6 'a t ~ mp ~ n ~ chevee. L'~ pld ~ r ~ plu3 1 ~ ~ ot p ~ r ~ tlell ~ -ment r ~ con ~ tltu ~ t denP ~ s ~ u ~; -Joc ~ t present ~
., lt ~ ~ r ~ 3 ldche et un ~ ~ r ~ nl ~ t ~ or no ~ hlclu ~ c de., c e11 ~ n ~ cl ~ os ~ s.
L '~ ocidtton ~ SF + dextrun ~ ulf ~ t ~ pr ~ nt ~
: t ca ~ .r, rn ~ r pui.; ~ ta ~, r ~ nt c ~ trls ~ nt In I ~ cc; ~ l ~ r ~ n-. ~ 'une ~, 3r; ~ r, ~ r ~ sus nutllr ~ l ~ rt ~ r ~ tlf, ~ t per. ~ -3 ~ t4nt ~ 'outr ~ p ~ rt a qur31Lt ~ increased ~ d ~ 1 ~ C cotrl ~ otlon p ~ r , s ~ n- a; 3- ~ all F, h ~ n ~ m ~ n ~ de tr ~ trac ~ l I ~ b unq It ~ obili: ~ atlon r ~ pld ~ -J ~, dlffér ~ nte3 c ~ t ~ orl ~ sc ~ liull ~ lr ~ :, n ~ -c; ~.;, 1r ~ ~ la r ~ st ~ ur ~ t'on ti ~ sul ~ lr ~.
P. ~ ur mie ~ 1 ll ust r ~ r 1 obJ ~ 2t o prb ~ ent o irr ~ nt, ~ n,, ~ n ~ en ~ cl ~ lr ~ m ~ l ~ t ~ n4nt u, 21q ~ n) pl ~ e:.
l: r ~ tL; ~ .rl ~ Ji r: e siont en ~ uc ~ no f4 ~; i ~ from $ tlnS3 ~ ~ n llrnltsr 1 ~ p ~ r; ~.
; ~ .'i G, '. ~ "R ~ ipl ~ r ~ p ~ ra ~ on p' ~ cr em ~
~ n pr ~ por ~ la composit ~ on su v-nte:
F ~ F.,., ................, ..... .. ,, .. ,, .., 10) I ~, ~ O! :: ax ~ r4ne ~ ul ~ t ~ ~ n 'P5660. ~ Uc ~ 4 42 ~ u ~ 51 ~ ns). ~ O m ~
C ~ rl ~ oxy ~, ~ th) ~ lc ~ lo ~ .. ............. ''. 5 ~, ~
ED U PUr 1fi ~: st ~ r11 * ~ t ~ pyr ~ n ~ a ...... 1 ~ ml ~ n ot ~ tlent. ~ insl un ~ ~ r ~ ma qe 1 'on ~ ppllqu ~ rT ~ rl ~ lt) ,. ct ~ iolr ~, ur ur ~ 3 pl ~ l ~ dq Syp ~: sci ~ r ~ Lc tLon, ou un r4t. ~ n t; ~ ur.e ~ trl ~, 3tlc. ~ n ~ pr ~ 3 3 ~ YES ~ c ~ traltornl ~ nt.

`:. , ',,' ~ ,,. ',;,. , ',',. .,. ,, ': ~,.

RCV B ': Xerox T ~ l ~ copi ~ r 7 ~ 20; 10- 4-as, 11:52 .33 1 ~ 74 51 37 ~ C., S ~ H: # 3 r 2; 1;

6. ~
r ,, ~! ~ r ~ r. ? P ~ ~ .p. ~, R ~ Jr ~ n ~ m. ~
~ ~ s ~ pp ~ rt ue ~ c, t un fll ~ coll ~ ne ~: ~. ~ ti .: te rlU ~ S (i ~ S. ~ cl ~ t ~ "F ~ t ~ C ~ L ~ n) t; ~ rllm ~ t, i: n ~ rC3rl ~ par ~ dsorptlerl pa ~ s ~ v ~ 3 du m ~ ll ~ se FGF-d ~ xtr; ~ ne ! i 3ulf ~ t ~ w dPwn ~ prepol tlons s ~ lv ~ n ~
Fr, F .... ~ ,,,,, ......................., .. 10, u, ~ e,; tran ~ ~ ulfat ~ ~ n- ~ 5 ~ 50 ~ u catal 3aSu ~ 51zm ~ O
~ al ~ 2. 1 ~ J. ~ l de s ~ ru ~ physlologlqu ~ ~ t ~ lo ~ t ~ pyr ~ g ~
r ~ fblt lr ~ cuber p ~ n ~ nt 3 ~ ~ lnute.a ~ 4 ~ CIe p ~ n ~ nt obtained, d ~, eupabl ~ ~ 'a forn ~ e ~ la pl ~ t nsu1, t ~ a sc ~ eu ~
v ~ d. ~ bundled.
It p ~ t ~ Sr ~ elppl1q ~ a. ~ Ce: ~ 'yl ~ r4 ~ ion ~
L, rOlur ~ 3s, d * c ~ upure. ~ And plal ~, prol ~ nd ~.
F. ~ t, ~ l, r. ~~ r ~ nf ll ~ ~
L ~ rr. ~ E pr ~ par ~ t lon qu ~ cl- ~ s $ u ~, 41n ~: oua le tnr5; n ~ me:! ~ op ~ roire, p ~ u ~ nt serve ~ impr ~ win utur ~ c ~ lrur31ce ~ 1 ~ t ~; F ~ ~ ehlcon - V1cl yl E ~ VZ). L ~ fll.
~ 7 iutrul-L ot ~ held e: jt used ~ bl ~ ~ n chtru ~ ~ o.

15l ~ lE, 4: Fl ~ r ~ tl ~ n,, ~ 'un .411 ~ r ~
We pr ~ p ~ r ~ e 13 compos' ~ I ~ n su vunte;
Y ~. ~ `..,. ,,,,,. ,,,,,,,,,,,,, .............. ~ 0 r.3 D ~: ~ tran ~ 6ulfi ~ t ~ ~ n 'D5650 cu C ~ ltA c13ue S ~ .sm ~). 150 L ~ rrn ~ 3 ~ rtlfici ~ oe .......... ............. 3 ml 011 ~ ff ~ ctu ~ In rn ~ l ~ nge p ~ r ~ lyophlll ~ tiorl, pu ~ n 4Jol ~ tr: 3 ml of e ~ u purl ~ i ~ e: ~ t ~ llo apyr ~ or cnt d ~ I'em7p1ci.
The coll. ~ Ri3 a.;. T 4ppliqu ~ ~ nx lcts c ~ $ d 'ul ~ ær, ~ L ion ~ -7 1 ~ l .orn ~ e, 4 rals, ~ n de ~ euY. ln ~ tlll tl ~ n $ p ~ r ~ our.

'~ 5

Claims (16)

1 - Composition including the association of minus an active form of a growth factor of the FGF family and at least one stabilizing dextran sulfate the biological activity of FGF. 2 - Composition according to claim 1, characterized by the fact that the dextran sulfate used is a water-soluble dextran sulfate, a dextran sulfate insoluble in aqueous medium, or a mixture of the two. 3 - Composition according to claim 2, containing at least one water-soluble dextran sulfate, characterized by the fact that the sulfated dextran (s) are present at a concentration of 0.1 µg to 50 mg per milliliter of final mixture. 4 - Composition according to claim 2, contain at least one medium-insoluble dextran sulfate aqueous, characterized in that the dextran (s) sulfates are present at a concentration of 0.1 µg at 500 mg per milliliter or gram of final mixture. 5 - Composition according to one of claims 1 to 4, characterized in that it contains from 0.01 ng to 300 µg of at least one active form of FGF, per milliliter or gram of final FGF / dextran sulfate mixture. 6 - Composition according to one of claims 1 to 5, characterized in that the FGF is a purified FGF
in its acidic or basic forms, or a mixture of the two, or still a derivative or analog of at least one of these shapes. 7 - Composition according to one of claims 1 to 6, characterized by the fact that the FGF / dextran association sulfate is incorporated into physiologically supports acceptable for in vivo applications. 8 - Composition according to claim 7, characterized by the fact that it constitutes a formulation aqueous, containing buffers and salts allowing maintain the FGF / dextran sulfate association in the vicinity physiological pH and in the vicinity of ionic strength physiological. 9 - Composition according to one of claims 7 and 8, characterized in that it contains additives, such as local anesthetics, substances bacteriostatic, sanitizing products, serum, or a water-soluble collagen. 10 - Composition according to one of claims 8 and 9, characterized in that it is placed under a directly usable form for application on the skin, as in solution, aerosol, anointing, lotion, powder, gel. 11 - Composition according to one of claims 8 and 9, characterized in that it is included in a pharmaceutically or cosmetically acceptable carrier, such than an ointment, a cream, a collagen gel, a gel of a other nature, or gelatin; that it permeates a gel of collagen, a gel of another nature, or another substance acting as a sponge, a dressing, a support covering bandage, or a biomateriau in front directly or indirectly promote repair tissue, such as a surgical suture; where she is applied to natural or synthetic fibers. 12 - Composition according to one of claims 8 and 9, characterized in that it is presented under a form applicable to the cornea, eye drops or gel intended to form a protective screen for the cornea. 13 - Composition according to one of claims 8 and 9, characterized in that it is included in liposomes. 14 - Composition according to one of claims 8 and 9, characterized in that it is presented under a locally applicable form internally on tissues and organs of the body, for example as a gastro-plaster enteric or lyophilized powders, whether or not included in capsules. 15 - Use of the composition as defined in one of claims 1 to 14, as an agent therapeutic, healing and tissue regeneration. 16 - Use of the composition as defined in one of claims 1 to 14, as an agent cosmetic, to improve the condition of the skin.