NGHIEN CU'U QUY TRJNH SU DUNG T^ BAG G 6 C D^ DI^U TR! MOT S6 BENH B^ MAT NHAN CAU
N g u y i n Thj Binh, Nguyen Manh Hi Trudng Dai hgc Y Hi N^i Nghidn cwu nhim xiy dwng quy trinh nuoi tao va ghdp ti/ thin tim bidu md nudi ciy tu' tS bao g6e bi4u mo vung ria, te bio goc bidu md nidm mpc midng vi td bio goc ket mac de dtSu tn hpi ehdng suy giim ^ bao goe vung ria. Ket qua da nudi tao thanh edng cac loai tam bieu md tir 3 loat ti bio gSc, cac tam biiu md cd eiu tmc hinh thai va hda hpc giong bieu md truoc giac mae, da ghdp thanh cong tim bieu mo nuoi eiy tir ti bio goc viing ria vd ti bao goc bieu md niem mac midng cho bgnh nhan bi hgi chimg suy giam te bio gde vting ria vdi ty Id 60 - 80%.
Tu' khoa: te bao goc, tam bieu mo, ghep md
I. DAT
V A NDt
Be mat nhdn c i u (BMNC) bj ton thuang do nhieu nguyen nhdn khac nhau; bdng m i t do hod chat hoge nhiet, hdi chirng Stevens- Johnson, n h i l m k h u i n mat nang, n h i l u gidi phdu thue hien tren ciing mdt m i t , deo kinh t i l p xuc... Mpt trong nhO'ng di chirng t h u d n g gap khi ton thuang be mat nhdn cau Id hpi chirng suy giam t l bdo goc ciia bieu md gide mac. Hau qua eiia hpi ehirng nay Id lam m i t dp trong cua giac mac do mang xa mach tir phia k i t mac xam l i n qua vung ria len b l mat gide mge, loet bieu md gidc mac khd han gdn, bong trdc bieu md giac mac tdi phdt. Vi vdy, thj lue m i t bj bgnh bj suy giam d n h i l u mire dp khde [ 1 , 2 ; 3; 4].
D l d i l u trj hdi ehirng suy giam t l bao g l c b i l u md gidc mac, eae nha nhan khoa da sir dyng n h i l u phuang phdp khde nhau nhu:
ghep mang oi dan thudn, ghep k i t mac ria t u thdn hoge dj thdn, ghep gide eung mge vimg ria. Vdi phuang phap ghep k i t mac ria, ghep
Dja chilien h§: Nguyen Thi Binh, bo mon M6 Phoi, truong Dai hgc YHa Ndi
Emait nauventhibinh-mo^vahoo.com vn Ngay nhan: 20n/2014
Ngay duoc chip thuin: 16/9/2014
gidc - cung mac ria t y than, manh md diing d l ghep d u g e lay tir mdt Idnh phai kha Idn. Ndm 1997, Pellegrini idn dau tidn dd sir dyng phuang phap ghep t i m b i l u mo giac mac d u g e nudi c i y tren mdng l i dh d i l u trj hOi chirng suy giam t l bao g l c viing ria gide mac tren ngudi [5].
Trong p h u a n g phdp nay, t l bao goe bilu md vimg ria d u g e l i y tir mat ben lanh hodc tir viing ria edn binh t h u d n g eiia m i t ton t h u a n g . Manh vung ria d u g e nuoi cay tren mang oi de tgo thdnh tam b i l u md giac mac.
Sau khi phdu tich h i t t l chire x a mgeh khdng binh t h u d n g tren be mat giac mae vd vimg ria, t i m b i l u md se d u a e co djnh tren b l mgt giac mge. P h u a n g phap ghep tam b i l u m l gidc mac nudi e i y d u g e cho la cd nhieu uu d i l m v u g t trdi so vdi cac p h u a n g phdp sir dung t r u d c day v i : mdnh mo smh thiet c6 kich t h u d e nhd, vige sinh t h i l t vung ria c6 t h i nhde Igi n i u e I n t h i l t , giam nguy ea thdi logi manh ghep vi trong t i m b i l u md gidc mae nudi eay khdng cd mat cua t l bdo Langerhans - te bdo ddng vai trd trinh dign khdng nguyen. Tuy nhien, p h u a n g phap ndy lai khdng thuc hien d u g e d ede t r u a n g hgp benh nhan ton t h u a n g ea hai m i t .
TCNCYH 90 (5)-2014
Doi vd'i cdc trudng hgp bdnh nhdn bj ton thuang be mat nhan cau ca hai mdt, tnrac ddy, ede bae sy nhan khoa da sir dyng phdu thugt ghdp vimg ria dj than, ghep k i t mge - ria dj than hodc ghdp tam b i l u md gidc mge nudi c i y di than. Tuy nhien, nhung benh nhan ndy phdi diing thuoe chong thdi logi manh ghep vd nguy ca thdi mdnh ghep kha eao.
Nam 2004, Nakamura va edng s u da nudi c i y thdnh edng t i m b i l u md tir t l bdo goc bieu md niem mac migng vd ghep t y than eho thd bl bdng gidc mac. M u d i ngdy sau ghep, tam bieu md g i n ehgt v d i md n i n eiia giac mac, Idp t l bdo tren ciing van edn nhdn vd khong ed hidn t u g n g sCrng hda [6] Leonard P.K. vd edng s y (2006) da nudi c i y t l bdo b i l u md nidm mac migng tren n i n mdng oi ngudi ciia 10 bgnh nhdn bj t h i l u hyt hodn toan vung ria, k i t qud la da tao ra dirge ede t i m bieu md g i l n g nhau ve mat hinh thdi. Sau khi ghep t u than eho bgnh nhan, chi sau 2 - 5 ngay da bieu md hod hoan toan b l mat gidc mge, 19 thdng sau ghep, be mat gide mac v l n I n djnh, thi lyc cua 9 trong 10 m i t tang gap hai lan [7]. Mdt so tac gid tren t h i gidi da nudi tgo t i m b i l u md tir te bdo g l c k i t mae de thay t h i b l mgt giac mge trong nhirng trudng hgp t i n t h u a n g be mat nhan c l u ca hai m i t . Nhu vay, vige su' dyng t i m b i l u md gidc mac nudi eay, tam bieu md niem mae mieng nuoi cay vd tam b i l u md k i t mge nudi c i y d l d i l u trj hpi ehung suy giam t l bao goc vung ria gidc mge la mpt l y a chpn tot cho cae benh nhdn bj ton t h u a n g b l mat nhan c l u mdt ben hoge hai ben. V d i p h u a n g phap nay, n g u d i bgnh khong phdi diing t h u l e ehong thdi logi manh ghep.
Vdi mong m u l n d u a vdo Vigt Nam mdt phuang phdp d i l u tri mdi cho eae bgnh nhan bj hpi chirng suy giam te bdo g l c vimg ria, ehung tdi t i l n hanh nghien ciru quy trinh s u dyng t l bdo gde de dieu trj mgt s l benh b l mgt nhdn cau nhdm myc tieu:
1. Xay dung quy trinh tgo tam b i l u md gide mge, tam bieu md niem mac mi#ng va t i m bieu md k i t mge tir te bdo goc viing ria giac mge, t l bdo gde b i l u md niem mge midng vd t l bdo goe bieu md k i t mge.
2. Xdy dung tieu c h u i n c h i t lugng ciia t i m b i l u md gidc mac, t i m b i l u md niem mge mieng vd t i m b i l u md k i t mac nudi c i y .
3. Chi dinh va xay dung quy trinh c i y ghep eae t i m b i l u md nuoi cay d l dieu tri t i n t h u a n g b l mat nhan edu
II. 0 6 l TU'ONG VA PHU'ONG PHAP
1 . D o i t u v n g
Nguyen bao sgi 3T3 do bg mdn Te bdo - Md Phdi va Ly sinh, Tardng Dgi hpc Khca hpe T u nhien eung edp.
Tho ehiing Orytolagus Cunieulus khoe mgnh, trpng lugng 2kg ± 0,2kg do trung tdm nghien ciru de vd thd San Tdy cung c i p .
Vung ria giac mac Idnh ciia benh nhan bi hdi chirng suy gidm t l bdo g l c viing ria mdt ben m i t : 10 benh nhan.
Niem mge mieng cua benh nhan bj hpi chirng suy giam t l bdo goc vimg ria hai m i t : 17 bdnh nhan.
K i t mge eua benh nhan t y nguygn: 6 benh nhan.
2. Md hinh nghien cCru
( 3 ^
( Nghien cuu m6i V irifcnq phu hffF
nch thCi V3 ):u' ly manh mg, ^ . ,
3. Cac bu'd'c nghien cuu
Nghien ciru t i l n hanh qua 2 giai dogn:
2.1. Nghien ciru thuv nghiem tren tho - Lua chpn vj tri sinh t h i l t vd kich t h u d e manh md nudi e i y :
+ Manh niem mge mieng: lua chgn giira 3 vi tri. viing giii'a ma, mat trong mdi, niem mac ma each mep 2cm.
+ Manh k i t mge: lua chpn giii'a 2 vj tri: k i t mac ciing d l va k i t mac nhan c i u .
Lua chpn mdi trudng nudi e i y SHEM1 hoacSHEM2.
L y a chpn phuang phap nudi c i y : nuoi b l n g manh md, nudi b l n g djch treo hoac nudi bang mdnh b i l u md.
Djnh danh t l bao cua tam b i l u mo nudi e i y bang eae ky thudt h i l n vi, hda md va hda md m i l n djch
Ghep t i m bieu md nudi edy eho thd bj gay bong gide mac. Theo doi va danh gia k i t qua ghep qua cdc b i l u hien lam sang va c l u triie VI t h i ciia m i t thd theo cac giai dogn: 7 ngdy.
14 ngay, 30 ngay, 60 ngdy, 180 ngdy sau ghep.
3.2. Nghien cii'u tren benh nhan tinh nguyen
- Sau khi cd k i t qua nghidn ciru trdn thyc nghiem, t i l n hanh nudi eay tam b i l u md v&
ghep t u than cho bdnh nhdn bj hdi ehirng suy gidm te bao g l c vimg ria mdt m i t ( t i m bilu md giae mae nudi e i y ) vd hai m i t ( t i m bieu md niem mge migng va tam b i l u md k i t mac nudi e i y ) .
Thdi gian nghien ciru: tir thdng 1 nam 2010 d i n thdng 6 nam 2013.
- Nai thye hien: Labo nudi eay md - Bp mon Md - Phdi, Tnj-dng Dai hpc Y Ha Npi, Khoa Kit - Gide mae, benh vign Mdt Trung u a n g .
4. Dgo difc nghien ciru
Nghien ciru da dirge Hdi dong dgo dire nghien eiru Y Sinh hpc, T r u d n g Dgi hpe Y Ha npi c h i p thudn. So g i i y chirng nhdn 77/HDDB - Y H N ngdy 1/7/2010.
III. K^T QUA
1. Nudi cay va ghep tam bieu mfi niem mac migng tren t h y c nghiem va tren benh nhan
1.1. Ket qua tren thin: n L y a chgn vi trf sinh t h i l t m d i ' e i y : niem mac vimg giira n
"'em tren thd'.
' o diing nuo h thude
76 TCNCYH &,
mdnh mo sinh t h i l t d l nudi d u g e 2 tam bieu md kich t h u d e 4cm^: 3 x 3 mm. Mdi tardng nudi c i y thich hgp: SHEM2 ( g i m DMEM/F12 ty le 1 : 1 , huyet thanh bdo thai bd (FBS) 10%, insulin, EGF, triiodothyronin, isoproterenoni vd hydrocortison, penicillin, streptomycin, amphotericin B, Phuang phdp nudi eay dan gian va cd higu qua cao: phuang phap nudi bang mdnh b i l u md vd sii" dyng nguydn bdp sgi t u thdn. Ty Id nudi tgo thdnh edng trong nghien eiru la 95,6%. Thdi gian nudi cay:
1 6 - 2 4 ngdy. Tam bieu md cd e i u true cua b i l u md lat tang khdng sirng hda, g i m 3 - 5 hang t l bao, cac t l bao lidn k i t vai nhau chat che bang cdc t h i lien k i t va cdc mdng bdo tuang, lien k i t ehgt vdi n i n mang oi b i n g cdc t h i bdn lien k i t . Cdc t l bao Idp trung gian cd K3 (+). Khdng ed eac t l bao t i l t nhdy xen vdo giCra cdc t l bao b i l u md tren cdc tieu ban nhudm PAS ( h i n h l , 2). D d nudi duge 79 t i m bieu md niem mge migng. Khi ghep t u than cde tam b i l u md niem mac mieng nudi c i y cho 15 thd bj gdy bdng gidc mac, 13 thd ed k i t qua tot (gidc mge trong, t i m bleu md lien t i t , nhan bdng, khdng edn tan mach, d mire dp vi the, cae t i m b i l u md dan sat vao nhu md giae va song tot), 1 thd k i t qua trung blnh, 1 thd cd k i t qua xau.
1.2. Ket qui tren benh nhan Dua tren k i t qua nghien ciru thuc nghidm tren thd, chung tdi da t i l n hanh nudi c i y 30 lan niem mge mieng ciia 17 benh nhdn (4 benh nhan nudi 2 l i n ) . Cd 3 benh nhan nudi e i y khdng thanh cdng. Sd t i m bieu md nuoi duge Id 54 (ty Ig nudi thanh edng la 90%. S l t i m b i l u md d u g e ghep Igi eho benh nhdn la 22. Nghien ciru dd thuc hign tren 22 p h l u thugt, d 19 mat ciia 14 benh nhan ed roi logn nang be mgt nhan c i u 2 m i t . Thdi gian theo ddi trung binh la 12,25 thang, tardng hgp dai n h i t Id 48 thdng, n g i n n h i t Id 2 thang. Phau
thudt ghep thdnh cdng d 12 ca: be mat nhdn cau on djnh, bieu md gidc mac nhan phdng, het cdc d i u hieu kieh thich c a nang. 9 ea thj lye edi thign (hinh 3). 4 trudng hgp t h i t bai do mdt b! ton t h u a n g nang. 4 trudng hgp t h i t bai do t i m b i l u md bj rdeh trong khi tdch d thi p h i u thuat.
2. Nudi cay va ghep tam bieu mo k i t mac tren thuc nghiem va tren b§nh nhan
2.1. Ket qua tren thtFc nghiem
Lua chgn vj tri sinh t h i l t m i u md dung nudi e i y : k i t mac cung d l . Kieh thude manh md sinh thilt: 0,5 x 6 mm. Mdi trudng nudi c i y thieh hgp: SHEM1 (gom DMEM/F12 ty le 1.1, huyet thanh bao thai bd (FBS) 10%, insulin, EGF, penieillin, streptomycin, amphotericin B.
Phuang phap nudi cay: phuang phdp nudi bang manh md, khdng sir dyng 3T3. Ty te nudi tao thdnh cdng trong toan nghien ciru la 8 6 , 1 % . Mdt sd tdm ed ldn nguyen bdo sgi.
Thdi gian nudi e i y : 1 6 - 2 4 ngdy. T i m bieu md cd e i u true ciia b i l u md lat t i n g khdng sirng hda, gdm 3 - 5 hdng te bdo, cdc te bao lien k i t vdi nhau chat che bdng cac t h i lien k i t vd eac mdng bao t u a n g , lien k i t chat vdi nen mdng di bang ede the bdn lien k i t . Cdc te bao Idp trung gian ed K4, K13 (+). Khdng cd cdc te bdo tiet nhdy xen vdo giira cdc t l bdo bieu md tren eac tieu ban nhudm P.A.S. Da nudi tao d u g e 54 t i m bieu md k i t mge. Khi ghep t y than cac t i m b i l u md k i t mac nudi eay cho 14 tho bi gay bdng giae mac, k i t qua:
2 mat ed k i t qua phdu thudt loai tot, 2 m i t k i t qua trung binh, 10 mat k i t qua xau.
2.2. Ket qua nghien cu'u tren benh nhan
Dua tren k i t qua nghien ciru thuc nghiem tren thd, chimg toi da t i l n hdnh nudi c i y 30 m i u k i t mac ciia 6 benh nhan. So t i m b i l u md nudi duge Id 20.
TCNCYH 90 (5) - 2014
Do k i t qud ghep tren thd khdng t i t , chimg tdi da khdng t i l n hdnh ghep cho benh nhdn.
2.3. Nuoi cay tam bieu md giac mac tip ti bao goc vung ria giac mac va ghep tip than cho binh nhan bj hpi ehu'ng suy giam ti bao goc vung ria mpt ben mat
Vj tri sinh t h i l t m i u md dimg nudi cay:
vimg ria gidc mge d vj tri 11 den 1 gid. Kich thud'c mdnh md sinh t h i l t de nudi d u g e 2 t i m b i l u md kieh thude 4cm^; 1 x 3 mm. Mdi trudng nudi cay thich hgp. SHEM1. Ty Ig nudi tgo thanh edng trong todn nghidn ciru la 100%. Thdi gian nudi eiy: 16 - 24 ngdy. T i m bieu mo co c l u true eiia bieu md Idt t i n g khdng sirng hda, g i m 3 - 5 hang t l bao, ede te bdo lien k i t vdi nhau ehgt ehe bdng eae t h i
lien k i t vd ede mgng bao t u a n g , lien k i t eh|l vdi nen mdng l i bdng cdc t h i bdn l i i n kit.
Cdc t l bdo lo'p trung gian cd K3, K12 (+).
Khdng ed cdc te bdo t i l t nhdy xen vdo gm cdc te bdo bieu md tren cdc tieu bdn nhuflm PAS (hinh 4). Chiing tdi da sinh t i i e t 10 manh vimg ria eua 10 bgnh nhan, nuoi 34 gilng, thu dirge 34 t i m b i l u md, ghep Igi cho b$nh nhk 10 tam, cdn 24 tam Idm ede ky thugt d l djnh danh t l bdo. Trong so 10 bgnh nhdn duge ghSp t i m b i l u md gide mge nudi e i y , ed: 6 bgnh nh9n cd k i t qud tot (hlnh 5), 3 bienh nhdn cd k i t quS tmng binh, 1 benh nhan ed k i t qua x i u . Thoi gian theo ddi trung blnh la 18,5 thang. Thj lire sau ghep tang tir 1 d i n 3 ddng d' cae bgnh nhdn ed k i t qua t i t vd trung binh.
Hinh 1 . Be mat tam bieu mo Hinh 2. Lien ket giira 2 te bao bieu mo
Hinh 3. Mdt bgnh nhan trud'c (A) va sau ghep t i m BM niem mge mieng 60 nqdv (B)
TCNCYH S
U5}-20li
Hinh 5. Mat benh nhan s a u ghep 1 nam K i t qua test dp te bdo trdn m i t cd k i t qua phdu thudt toL
IV. BAN LUAN
Phuang phdp ghep t u than tdm bleu md nudi c i y tir cdc nguon t l bdo goc khde nhau de d i l u tri hoi ehung suy giam t l bdo goe vimg ria hodn todn mdi doi vdi Viet Nam. Vdi phuang phap nay, bgnh nhdn khdng phdi sir dyng thuoc ue e h i m i l n djch. P h i u thuat ghep t i m b i l u md nudi eay cd dd an toan cao, ed the Igp Igi n h i l u l i n md khong dnh h u d n g d i n tinh trgng m i t eung n h u todn than cua ngudi bdnh. Khi p h l u thudt ldn t i l p theo, chi can ggt bd lo'p xa mach tren b l mat gidc mac va thay the bdng tam bieu md nudi e i y , do dd khdng Idm ton hgi gi them eho to chu'c nhu md d phia d u d i . V l n d l quan trgng n h i t cua phuang phdp ndy Id nudi tao thanh cdng trong
phdng thi nghiem t i m b i l u md tir cdc nguon t l bao g l c khac nhau. Cde yeu t l q u y i t djnh s u thdnh edng eiia qua trinh nuoi cay Id: vj tri l i y t l bdo goe, mdi trudng sir dung nudi cay, phuang phdp nudi cay. Trong nghidn eiru, ehung tdi dd xdc djnh d u g e vj tri lay t l bdo g l c d' vung ria Id vj tri tir 11 - 1 gid, d niem mac mieng Id mgt trong vimg trung tam ciia ma, d k i t mac Id k i t mac ciing d l . Chung toi ciing xac dinh d u g e mdi trudng nudi cay thich hgp d l i vdi t l bdo goe vimg ria va k i t mac la mdi trudng SHEM1, edn vdi t l bao gde bieu md niem mac mieng la mdi trudng SHEM2.
Hign nay, tren T h i gidi thudng sir dung 2 phuang phdp nudi edy: nuoi b l n g manh mo vd nudi bang djch treo. Nghien eiru eua chimg
tdi eho t h i y : khi nudi e i y te bao g l c vimg ria vd te bdo g l c k i t mge, sir dyng phuang phdp nudi bang manh md dan gian vd cd hieu qud.
D l i vdi vige nudi e i y t l bdo gde bieu md niem mae migng, chiing toi dd tim ra mot phuang phdp nudi e i y hodn toan mdi, chiing tdi gpi Id phuang phdp nudi c i y bdng manh b i l u mo.
Vdi phuang phdp ndy, k i d i thude manh md trich thii nhd (3 x 3mm), phuang phdp xir \-y mdnh md dan gidn, khdng sir dung sdn p h i m cua dpng vdt (Idp 3T3), tgo dieu kien eho cde t l bdo b i l u md phdt tnen tren md n i n giong trong nidm mac mieng, ty le nudi tao thanh cdng t i m b i l u md eao
C h i t luang cac logi tam bieu md nudi c i y cua Chung tdi tuang duang vdi cdc tde gia trdn t h i gidi vd da duge vien K i l m nghiem Q u i e gia cdng nhdn. Tuy nhien, d l i vdi t i m b i l u md nudi c i y tir t l bao g l c k i t mac, chat lugng cdn chua on djnh, mdt s l cdn l l n nguydn bdo sgi. Cac t i m b i l u md nudi c i y deu duge ghep vd danh gid k i t qud ghep tren thuc nghigm. K i t qud ghep tren thye nghiem cho t h i y tam bieu md nuoi tgo tu" t l bao g l c viing ria vd b i l u md niem mac mieng bam dfnh va ton tai tot trdn mat thd b| bdng, edn ghep t i m bieu md nuoi eay tir t l bao goc k i t mae cho k i t qud khdng t i t va chimg tdi da khdng ghep t h u nghiem tren benh nhan. Vdi cde benh nhdn d u g e ghep t i m b i l u md nudi tgo tir t l bdo goc vung ria hoac t l bdo g l c b i l u md niem mae mieng: mdng l i ap t i t len b l mat Idp nhu md gidc mge, giae mac trong, n h i n bdng, hien tugng viem dwge k i l m sodt, trigu ehirng cgm chdi, ehdy nude mat, khd chju deu giam vd m i t di sau phau thuat, t h j ' lye sau mo tdng tu" 1 den 3 ddng so vdi thj lye trudc phau thuat.
V. KI^T LUAN
Da xdy d y n g thdnh edng qui trinh nudi tgo
t i m bieu md gidc mac tir t l bao goe viing ria gide mge, tam b i l u md nidm mae migng tit t l bdo goc bieu md niem mac mieng. D l i vdi qui trinh nudi tao t i m b i l u md k i t mac ttr t l bdo g l c k i t mac: dd nudi tgo thanh edng tam b i l u md k i t mae, tuy nhien chua xdc djnh d u g e do tinh k h i l t (lan nguyen bdo sgi) cua tam b i l u m d .
Da dua ra vd da dirge k i l m djnh tieu ehuin ea s d eua cdc t i m b i l u md nudi eay; V6 k h u i n , rdng 4em^, Id bieu md Idt t i n g gom 4 - 5 hdng t l bao, ede t l bdo g i n ehgt vdi nhau bang mdng bdo t u a n g va t h i lien ket, g i n vdl mang oi bang the bdn lidn k i t , bao t u a n g cd ede keratin dae higu.
Da xdy d u n g d u g e qui trinh ghdp tim bieu mo gide mge nudi cay vd t i m bieu md niem mae midng nudi e i y d l d i l u trj b|nh nhSn bj t i n t h u a n g b l mat nhan e i u mdt mat vd hai m a t
L o ' i c a m a n
Chung tdi xm tran trpng cam a n Bd Khoa hoe va Cdng nghe da cap kinh phi cho chung tdi thuc hien de tai dge lap cap Nha nude nay Chimg toi cam k i t ddy Id ket eua nghien ei>u eua nhdm chiing toi, khdng trung lap vdi nghien ciru nao khac.
TAI LIEU THAM K H A O
1. A b r a h a m S (2009). Ocular surface reconstruction. From tissue transplantation to cell therapy. Tech Ophthalmology, 7,118 -123.
2. Shortt A . J . , Seeker G.A., Notara M.D et al (2007). Transplantation of Ex Vivo Cul- tured LImbal Epithelial Stem Cells. A Review of Techniques and Clinical Results. Surv Oph- thalmol. 52(2), 483 - 502.
3. A l f a n o C , C h i u m m a r i e l l o S., F i o r a m o n t i P et al (2006). Ultrastructural study of autologous cultivated conjunctival
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epithelium. Ophthalmic surgery lasers and Imaging, 37(5), 378 - 382.
4. Grueterich M., Espana E.M., Touhami A et al (2002). Phenotypic study of a ease with successful transplantation of ex vivo expanded human limbal epithelium for unilat- eral total limbal stem cell deficiency. Ophthal- mology, 109(8), 1547 - 1 5 5 2 .
5. Pellegrini G., Traverco C.E., Franzi A.T et al (1997). Long-term restoration of damaged corneal surfaces with autologus cultivated
comeal epithelium. Lancet, 349, 990 - 993.
6. Nakamura T., Inatomi T., Sotozono C et ai (2004). Transplantation of cultivated autologous oral mucosal epithelial cells In patients with severe ocular surface disorders.
Br J Ophthalmol, 88, 1280-1284.
I. Leonard P. K. Ang, Takahiro Naka- mura, Tsutomu Inatomi et al (2006). Autolo- gous Serum - Derived Cultivated Oral Epithe- lial Transplants for Severe Ocular Surface Disease Arch Ophthalmol, 124, 1543 - 1 5 5 1 .
Summary
OPTIMAL PROCEDURE OF STEM CELL THERAPY TO TREAT SEVERE OCULAR SURFACE DISORDERS
The purpose of this experiment is to determine the optimal modality to cultivate and transplant epithelial cell sheets from limbal stem cells, oral mucosal stem cells and conjunctival stem cells to treat Limbal stem cell deficiency (LSCD) symptoms. We succeeded in creating epithelial cell sheets from three different types of stem cells. Confluent sheets of epithelial cells at the harvest- ing day possess the morphological and immunohistoehemieal features of corneal epithelium.
The ocular surfaces were successfully reconstructed in 60 - 80% of LSCD patients with autolo- gous cultivated oral mueosal/limbal epithelial sheets.
Keywords: stem cells, epithelial cell sheets, transplantation
TCNCYH 90 (5) - 2014
