TAP CHI SINH HOC 2014, 36(1): 125-132 DOI: 10.i5625/0866-7i60-v36nl 4530'
BIET HOA TE BAO GOC TRUNG MO DAY RON THANH TE BAO GIONG TE BAO BliU MO DA
Doan Chinh Chung''^*, Le Thanh Long^, Hoang NghTa Son^, Nguyen Hoang Chuwng', D o Minh Si^, Le Van D o n g '
'Trudng Dai hgc Khoa hgc ty nhien, tp. Hd Chi Minh ' H o c vifin Quan Y, Hd Ndi
'vifin Sinh hpc nhifit ddi, Vien Han Idm K H & CN Viet Nam, *[email protected]
TOM TAT: Te bdo gfic (TBG) tmng mo day ron cd the thu nhan ma khong gap phai nhung rao can ve dao diic. Cdc tfi bao nay c6 kha nang tang sinh manh. ty lam mdi va tiem nang bifit hda thdnh nhieu loai te bdo khac nhau. VI vay, nghien ciiu nay tap ming danh gia tiem nang biet hda ciia TBG tmng mo day rdn thanh c^c te bao bieu mo da. Trong nghien ciiu nay. cac TBG iing vifin dupc phan lap bdng ky thuat nuoi cay mo bam dmh va dupc mo ta mdt sd dac diem ve kieu hinh miln dich, tiem nang biet hda. Te bao sau 3 lan cay chuyen dupc cam img biet hda thanh te bao bieu md da. Ket qua cho thay, cac TBG day rdn co hinh thai gidng nguyen bao soi, tiem nang tang sinh manh va biet hda thanh te bao md, nguyen bao xuong. Cac te bao nay duong tinh vdi CD73, CD90, CD105, CD166, nhimg am tinh vdi cac marker ciia tk bao mau CD19, CD34, CD45 va HLA-DR. Sau qua trinh biet hoa, cac tfi bao dupc cam ung cd sy thay doi hinh thai nhat dinh so vdi cac tfi bao doi chiing, dong thdi bieu hien mot sd gen Cylokeralin 18 (CK18), Cytokeratin 19 (CK19) va P63, trong khi cac tfi bao doi chiing khdng bieu hien cac gen nay. Dac biet, ca te bao bifit hda va te bao ddi chimg deu bieu hien gen fil -integrin. Nhiing ket qua nay chimg to TBG trung mo tu day rdn cd the biet hda thanh tfi bao bieu mo da in vitro dudi dieu kien nudi cay phti hpp vd c6 the su dyng nhu mot nguon te bao ly tudng cho ky thuat md da nhan %o.
Tir khoa: Bift hda, day rdn, te bao gdc tmng md, te bao bieu md da.
MOfDAU
Nhirng nam gan day, vific irng dyng lieu phap tfi bao gdc (TBG) dd ft nhifiu mang dfin nhiing thanh cdng nhat dinh trong viec phyc hdi cdc ton thuang Idn tren da do bong sdu hay khuyfit hdng da ldn [9, 14]. Hifin nay, ben canh ngudn TBG ty than tir tuy xuang, cdc TBG trung md giau tifim ndng tir mo day rdn cung dupc chu trpng nhifiu trong dieu tri cay ghep tu than va cdy ghep khdc gen ciing loai. Nguyfin nhan do cac tfi bao nay sd hiru mpt so uu diem vugt trpi nhu tinh thai loai bdi he thong mifin dich cay ghep thap, cd khd ndng lic ehfi vd difiu bien mifin dich, tifim nang tang sinh va biet hda cao, khdng gap phdi vdn dfi vfi y dao diic vd ngudn mau thu nhan ddi ddo [8, 20]. vi vay, cd thfi su dung de cay ghep cho cdc bfinh nhdn, ddc bifit trong trudng hpp khdng co san ngudn TBG ty thdn. Tuy nhifin, khi img dung TBG trung mo day rdn vao difiu trj cac tdn thuong da, van chua cd thdng tin vfi van dfi lieu cac tfi bao nay cd the bifit hda tryc tifip thanh cdc tfi bdo da hay khdng.
Nghien ciru cua Dai et al. (2007) [6] tren md hinh chupt bj tdn thuang khuyfit hdng da cho thdy, cac TBG tmng mo tir mdu ddy rdn cd the tdn tai trong 2 tuan xung quanh vi tri tdn thuong, va thiic ddy qua trinh lanh hda vfit thuang thdng qua khd ndng bifit hda thdnh cac tfi bao bifiu md duong tinh vdi CK87CK10"^ sau khi dupc cdy ghep. Ngoai ra, nghifin ciru ciing phat hifin thay cac te bdo duong tinh vdi N S T Y * / H L A - r trong cdc tfi bao dupc phan lap tir mau da sinh thifit. Tuy nhifin, nghien cim ciia Tark et al. (2010) [21] cho thiy, cac tfi bao nay thiJC day qua trinh lanh hda vet thuong thdng qua c a ehfi can tifit bang each gia tang ham lugng yeu td chuyen dang p (TGF-p), mpt trong cac yfiu to ddng vai trd quan trpng quyfit dinh kha nang lanh hda vet thuong, thay vi biet hoa thanh cac te bdo da. Hay cdc TBG trung mo tir mdng di co khd nang thiic ddy sy ldnh hda vfit thuang tren md hinh chupt dai thao dudng bj tdn thuong khuyfit hong da thdng qua hai ca che: bifit hda thanh mdt sd tfi bao bifiu md
Doan Chinh Chung etal.
duong tinh vdi CK19* trong viing md tdn thuong va kich thich qua trinh hinh thdnh mach mau mdi thdng qua su gia tang ndng dp mgt sd yfiu td nhu IGF-1, EGF va IL-8 [12]. Ngugc lai, nghifin ciiu cua Schneider et al. (2010) [19] cho thdy, TBG trung md tii day ron hd trg qua tnnh ldnh hda vfit thuang da bang kha nang biet hda thdnh cdc nguyfin bao ca vdi sy bieu hien ddng thdi cac marker bieu md va trung md pan- CK7SMA7vimentin*.
Mac dii cd rat nhifiu nghifin ciiu danh gia tiem nang biet hda in vivo cua cdc TBG trung md day rdn trong vific thiic day qua trinh lanh hda vfit thuong, tuy nhifin, cd rdt it cac nghifin cmi in vitro dugc bao cao ve van de nay.
Nghifin cuu cua Kamolz et ai. (2006) [11] cho thay, cac TBG tii mau dly rdn co thfi bifit hda thdnh cdc tfi bao bifiu mo in vilro khi dong nudi cay vdi cac tfi bdo bieu md trudng thdnh. Tuy nhien, nghifin ciiu khdng chi ra cy thfi Ioai TBG nao trong mdu day rdn (TBG tao mau hay TBG tmng md) thyc su tham gia vao qud trinh nay.
Nghifin cuu ciia Tran et al. (2011) [22] ciing cho thdy, tiem nang bifit hda thanh cdc te bao simg in vilro cua TBG tmng mo mau ddy rdn khi dugc nudi cay vdi mpt sd yeu td nhu EGF vd KGF. Viec thifiu mpt md hinh in vilro gay can trd cho vific nghien ciiu cac co ehfi phdn tu, cac yfiu td ddng vai trd quan trong trong sy dinh hudng bifit hoa thanh cac tfi bdo da va sy hinh thanh cau triic da hoan chinh trong cac cd the sau khi dupc cay ghep TBG. Vi vay, viec danh gid tiem nang bifit hda cua TBG tmng md tir day ron thanh te bao bifiu md da in vilro la mdt hudng di cdn thiet ciia vific iing dung TBG trong ky thuat md da nhan tao.
V^T LIEU VA PHirOfNG PHAP NGHIEN ClTU Phan lap va nudi cay TBG ling vifen tir day r6n
Mau day rdn (dm tinh vdi HBV, HCV, HIV) sau khi thu nhan dupc qudn bao trong dung dich PBS cd b6 sung 2% khang sinh Pen-Strep (GIBCO Invitrogen, Dire) d 4''C cho tdi khi thao tac. Mdu day rdn dugc nia lai 2-3 ldn vdi PBS va dupc cat thanh tiing doan khodng 2-4 cm. Cac doan nay dugc cdt thanh tirng tam nhd, tdch bo dpng mach, ttnh mach cua day rfin va
Idp dich nhay Wharton's Jelly dpc theo chifiu ddi ciia tam. Sau dd, cac doan ddy rdn dugc cat thanh cac mdnh md nhd vdi kich thudc 2-3 mm' vd dupc chuyfin vao dia nudi cay md 100mm.
Mdi frudng nudi cdy gfim DMEM/F12 (GIBCO Invitrogen), 15% FBS, lOng/ml FGF, 10 ng/ml EGF, 2 mM L-glutamine, IX insulin- transferrin-seienium (ITS) (Sigma-Aldrich, Hoa Ky). Cac man mo dugc nudi cay d 37"C, 5%
C02 va mdi trudng nudi cay dugc thay 2 lin/tudn. Khi tfi bao dat 70-80% di?n ti'ch be mat dimg cu nudi cdy, tien hanh cay chuyfin bdng Trypsin/EDTA 0,25% (Sigma-Aldrich) theo tyle 3:1.
Phan tich marker be mat cua TBG ihig vifin Cdc TBG ling vifin sau 3 lan cay chuyfin dugc xdc dinh su bifiu hifin cac marker bfi mat bang kl thuat flow cytometry. Khoang 10^ tfi bdo dugc sir dung de nhudm vdi khang the trong 30 phiit d 4''C. Sau dd, cac tfi bdo nay dugc rua lai 2 ldn bang dung dich FACSflow dfi loai bo khang the thira vd tai huyfin phii trong 500 pi dung dich FACSflow. Khi tifin hdnh phan tich, cdc khdng thfi cd gdn chat phdt huynh quang HTC hay PE: CD19-FITC, CD34-FITC, CD45- HTC, HLA-DR-FITC vd CD73-PE, CD90-PE, CD105-PE, CD166-PE (BD Biosciences, Hoa Ky) Idn lugt dupc phdt hien bdi kfinh FLl va FL2. Tat cd cac dd lieu dugc phdn tich bang phdn mem CellQuest Pro trfin he tiidng FACSCaUbur (BD Biosciences). Toan bg thi nghiem dupc lap Iai ft nhdt 2 ldn.
Bifit hda TBG irng vien thanh nguyen bao xirong va te bao md
Dfi biet hda thdnh tfi bdo md, cac tfi bao dupc cam iing bdng mdi tradng DMEM-low glucose, bd sung 10 pM dexamethazone, 10 pg/ml insulin, 200 pM mdomethacine, 0,5 mM Isobutyl-metylxanthane va 10% FBS (Sigma- Aldrich). Hieu qua bifit hda dupc ddnh gid thong qua quy trinh nhupm Oil Red [13]. Tfi bao dugc cd dinh bang dung dich paraformaldehyde 4%
trong 1 gid, sau do rira lai bdng isopropanol 75% va nhudm vdi thudc nhudm Oil red trong 10 phiit. Cudi ciing, tifin hanh khii nhudm bdng isopropanol 100% trong 15 phiit. Dfi biet hda thanh nguyfin bao xuong, cac tfi bao dugc cam ling bang mdi trudng DMEM-low glucose, bd
Biet hda te bao gdc trung md day rdn sung 10 mol/L dexamethasone, 50 pmoI/L
ascorbic acid-2 phosphate, 10 mmol/L p- glycerol phosphate va 10% FBS (Sigma- Aldrich). Ket qua biet hda dugc ddnh gia thdng qua phuong phdp RT-PCR [13]. Cac quy hinh biet hda dugc tien hanh trong 2 tuan. Te bdo dupc nudi cdy trong mdi trudng DMEM-low glucose, 10% FBS dugc sir dung lam ddi chiing.
Bifit hda TBG t r u i ^ mo thanh ti bao bi^u md da'
Quy hinh bifit hda dugc tifin hdnh trong 3 tudn gdm 2 budc. Budc I: te bao dugc nudi cay trong mdi tmdng DMEM-low glucose, bo sung 10 ng/ml KGF vd 20 ng/ml EGF (Sigma- Aldrich) trong 1 tudn ; budc 2: sau 1 tuan nudi cdy, tfi bao dugc chuyfin sang mdi tmdng SFM (GIBCO Invitrogen) cd bfi sung 20 ng/ml HGF va 50 ng/ml IGF-2 (Sigma-Aldrich) trong 2 tudn; te bdo ddi chimg dupc nudi cay trong DMEM-low glucose, bd sung 10% FBS. Sy thay ddi hinh thai cua te bdo ddi chiing va tfi bdo bifit hda dugc theo doi lifin tyc trong qua trinh bifit hda. Cdc te nay cung dugc ddnh gia sy bifiu hifin mgt sd gen ddc trung cua te bao bifiu mfi nhu CKIS, CK19, P63 vd ^1-inlergrin bdng ky tiiuat RT-PCR.
Thu nh^n RNA thn% so va RT-PCR RNA tong sd ciia tfi bao bifit hda va doi chirng dugc tdch bdng Trizol (Sigma-Aldrich) theo hudng dan cua nha sdn xuat. Phan irng RT- PCR dugc thuc hifin bdng bd kit AccessQuick™
RT-PCR (Promega, Hoa Ky) vdi phan iing phien ma ngupc: 45 phut d 45°C, kfit thuc d 95''C trong 2 phut. Chu trinh nhifit PCR: 94°C ttong 45 giay, 5 5 ^ - 6 0 ^ ttong 45 giay, 72''C trong 1 phiit vdi tdng sd chu ki 35. Sau khi chu ki CUOI hoan tdt, u 10 phiit d 72''C. Sdn pham PCR dugc phan tich dien di ttfin gel agarose 2%
va quan sdt bdng he thdng chup gel ty ddng GelDoc It (UVP, Hoa Ky). Trinh tu cac primer:
Osteopontin F: CTAGGCATCACCTGTGCCA TACC, R: CTACTTAGACTACrrGACCAGT GAC (330) Osteocalcin F: CGCAGCCACCGA GACACCAT, R: GGGCAAGGGCAAGGGGA AGA (405bp). CK18 F: TGGTACTCTCCTCA ATCTGCTG, R: CTCTGGATTGACTGTG GAAGT (149 bp). CKI9 F: AGGTGGATTCC GCTCCGGGC, R: ATCTTCCTGTCCC
TCGAGCA (460 bp). P63 F: CAGACTCAAT TTAGTGAG, R: AGCTCATGGTTGGGGC AC (611 bp). pl-integrin F: AATGTTTCAGTG CAGAGCC, R; TTGGGATGATGTCGGGA (640 bp). GAPDHhu: F: ACAGTCAGC CGCATCTTCT T. R: ACGACCAAATCCGTT GACTC(94bp)[4, 13, 22].
KET QUA VA THAO LU^N
Hinh thdi va kba nang tang sinh ciia TBG trung md
Su xuat hien cac te bao bdm ttdi cd hinh thai giong vdi nguyfin bdo sgi d giira cac khodng ttdng cua cdc khdi mo dugc quan sat thay sau 7 ngay nudi cay (hinh la). Sau 10-12 ngdy nudi cdy, cdc tfi bao bam ttdi hinh thdnh cdc cum tfi bdo xung quanh cdc khoi md. Vao thdi difim nay, cac khdi mo dugc loai bo. Cac tfi bao bam ttdi ddn dan dinh hudng theo mdt chifiu nhdt dinh. Sau 3 lan cdy chuyfin, cac tfi bao cd su ddng nhdt tuong ddi vfi mat hinh thai (hinh lb).
Te bao dupc phdn lap tir ddy rdn ttong nghifin cihi ndy cd hinh thai tuong tu nhu cdc TBG tmng md dugc phdn lap tir tuy xuong ngudi, tuy nhifin, kha nang tdng sinh ciia TBG tmng md tir day ron Idn hem so vdi TBG tmng md tu tuy xuang [3].
Tiem nang biet hda in vitro va kieu hinh mien djch ciia TBG trung md
TBG ttung mo dupc nudi cdy trong mdi tmdng thich hpp de cdm iing bifit hda thanh nguyfin bdo xuong. tfi bao tao md. Tfi bdo tao md dugc biet hda tir TBG tmng md cd hinh thai CO tton, cac gipt lipid xuat hifin, chifim phan ldn lfi bdo chat va bat mau do vdi thuoc nhudm Oil red (hinh Id), ngupc Iai, cac tfi bdo doi chimg khdng cd sy thay doi vfi mat hinh thdi (hinh Ic).
Tuong nr, cdc nguyfin bao xuang dupc bifit hda tir TBG trung mo bifiu hifin hai gen dac tnmg ciia tfi bdo xuong la osteopontin va osteocalcin sau 2 tudn bifit hoa. trong khi dd, cdc te bdo doi chimg khong bifiu hifin hai gen nay (hinh 3b).
Cac marker bfi mat ciia tfi bao sau 3 lan cdy chuyfin dugc phan tich bang he thong flow cytometer, cac marker duong tinh va dm tinh dupc phan lich dong thdi. Kfit qua cho thay, hau hfit tfi bao dfiu am tfnh vdi cdc marker ciia te bdo mau nhu CD19 (khang nguyen cua tfi bdo
127
Doan Chinh Chung etal mono), CD34 (marker dac trung cua TBG tao
mau), CD45 (khang nguyen cua te bdo bach cau trudng thanh), HLA-DR (bieu hien d te bdo mono, dai thuc bdo vd te bao lympho B) vd duong tinh vdi cac marker CD73, CD90, CD105 vd CD166 (hinh 2), ttong do CD73,
CD90 va CDI05 dugc xem nhu la cac marker quan ttgng de nhan dipn TBG trung mo ngudi [7]. Kfit qua nay chiing td quan thfi tfi bdo phan tt'eh khdng tap nhifim vdi cdc ddng te bdo man va cd kieu hinh nufin dich tuong ddi dong nhdt sau 3 Ian cay chuyen.
i''tfl ha
Hinh 1. Hinh thdi ciia ciia TBG tir day rdn sau 7 ngay nudi cdy (a) va sau 3 ldn cdy chuyfin (b). Sy thay ddi hinh thai tfi bdo trade (c) va sau khi bifit hda thdnh tfi bdo md, tfi bao bifit hda bdt mdu dd vdi thudc nhudm Oil Red (d). Tfi bao chuyen dan tir ttang thai bdm ttai (e) sang ttang thai co ttdn vd tang sinh chdm (f) sau qua trinh bifit hda thdnh te bao bilu md da.
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Hinh 2. Ket qua phan tich cac marker he mat cua TBG tir day r6n b3ng he th6ng mdy flow cytometer vcri cac khang thg co g4n chat phat huynh quang FITC hoac PE.
Biet hda te bao gdc tnmg md day rdn
( a ) , G"^H ^ fl-aHgrU^ CK19 ^ CKlg ^ ^ P63 ^ ( b ) GAPDH _ OWwuakin ^ ^ OrteoiwoU
Hinh 3. Ket qud phdn tich RT-PCR. Su bifiu hifin CKIS, CK19, pl-inlergrin va P63 cua tfi bdo trudc va sau khi bifit hda (a). Te bao ddi chiing (gieng I), te bdo biet hoa sau I mdn (gifing 2) va sau 3 tu4n (gifing 3). Sy bifiu hifin osteocalcin vd osteopontin ciia tfi bao tmdc va sau khi biet hoa (b).
Tfi bdo ddi chiing (gieng 1). Te bao biet hda sau 1 mdn (gifing 2) va sau 2 tudn (gifing 3). GAPDH sit dyng Idm ddi chiing ndi.
Nhu vay, cac TBG tmng md tii ddy rdn ttong nghifin ciiu nay cd cac ddc diem sinh hgc tuong ddng vdi kfit qua cua cdc nghifin ciiu tmdc dd [3,18, 23], ddng thdi ciing dap iing cac tifiu chl md Oy ban Te bao gdc phdi va tmng md ciia Hifip hgi Hfiu phdp Tfi bdo quoc tfi (ISCT) dua ra de xdc dinh TBG tmng md ngudi [7].
Kbd ndng bifit boa in vitro cua TBG trung md thdnh tl bdo bieu md da
Trong Id thi nghiem ddi chiing, tfi bao vdn duy tri ti^ng thai bdm trdi vd tang sinh manh sau 1 tuan (hinh le), trong khi dd, cac tfi bao cdm ling bifit hda cd su thay doi hinh thai ro rpt. Cdc te bdo nay chuyfin tir ttang thai bam trai sang ti?ng thai co ttdn va tang sinh chdm (hinh If)- Ngodi ra, trong qud trinh quan sdt su thay doi hinh thai ciia cac te bao bifit hda, nhom nghifin ciiu nhan thdy sy thay ddi hinh thai ciia tfi bao dudng nhu xay ra ro rdng hon va nhanh han d nh&ng viing cd mat dp te bdo cao so vdi nhirng viing CO mat dp tfi bao thap. Difiu nay cho thay, sy tuang tac giua cdc te bdo cd thfi Id mdt ttong cdc yeu to can thifit cho sy chuyen bifit hda cua ddng tfi bdo tmng md thanh ddng tfi bdo bifiu md [I]. Hon nira. trong nghifin ciru ndy, nhdm nghifin ciiu ciang tifin hdnh tach va Ioai bo Idp bifiu md, tinh mach va ddng mach day ron trade khi nudi cdy, do do, vific tap nhifim cac te bdo bifiu md trong nudi cay cd the tranh dupc. Nhu v^y, sy thay ddi hinh thai cua tfi bao sau qua trinh bifit hda tuong tu nhu cdc kfit qua nghien cuu da dupc cfing bd trade ddy khi tien hanh nudi cay cdc TBG tir mau day rdn trong mdi tmdng cd sy hifin dipn cua cdc yfiu to tang
tmdng nhu KGF va EGF [22] hay ddng nudi cdy vdi cdc te bdo simg [II].
Cac tfi bao bifit hda bifiu hien CKIS sau 1 tuan cdm irng bifit hda (sau budc I) va bifiu hifin CK19 va P63 sau 3 tudn cam ting bifit hda (sau budc 2) khi phan tich bdng ky thuat RT-PCR (hinh 3a). Trong dd, P63 cd thfi dupc sir dung nhu mdt marker dfi nhan dien te bdo bifiu md da hay tfi bao simg [16, 22]. CK18 va CK19 Id cac marker ddc trung cua cdc tfi bao tifin than bieu md hay TBG da [10, 15]. Sy thay dfii hinh tiidi cua tfi bao kfit hpp vdi sy bieu hien mdt so gen ddc tnmg cua tfi bao bifiu md chihig to sy chuyfin bifit hda cd xay ra trong tfi bdo bifit hda sau qud trinh cam iing. Kfit qud nay cang dugc Cling CO khi phan tich sy bifiu hifin cua ^1~
integrin. Dac biet, fil-integrin dupc phat hifin trong cd tfi bao ddi chihig va te bdo biel hda va su bifiu hifin cua fll-integrin trong tfi bdo bifit hda cao hen so vdi tfi bdo ddi chimg (hinh 3a).
Dieu ndy chiing to cac TBG trung md tir day ron cd tifim ndng biet hoa thdnh cac te bao bifiu md. Vi pl-integrin dupc xem nhu mpt marker trung gian chi thi cho su chuyen biel hda giiia Idp trang md va ldp bifiu mo [17]. Ngoai ra, mdt sd nghien ciru cho thay mirc dp bifiu hien ciia pl-integrin dnh hudng ldn den su bifit hoa thanh te bdo simg in vilro [2].
Nghien ciiu ciia chiing tdi chi dimg lai d muc dp danh gid su thay doi hinh thdi, sy bieu hifin mdt sd gen d muc phifin ma mRNA, chua tifin hdnh danh gia d miic dp protein vd cdc thu nghifim in vivo dfi xdc djnh cdc tfi bao biet hoa cd thuc sy Id cac te bao bieu md da tmdng
Doan Chinh Chung el al.
thanh hay khdng. Vi vay, chiing tdi gpi cac tfi bdo biet hda nay Id cac te bao gidng tfi bao bieu md da. Tuy nhien, kfit qua ciia n ^ e n ciiu nay ciing cho thdy, ttong q u a ttinh biet hda thanh dong tfi bao bifiu md, cac te bao tien than cd thfi xuat hien ttong giai doan dau cua qua trinh bifit hda (sau I tuan cdm iing, tfi bdo chi bifiu hien CKIS vd pl-intergrin) vd cdc tfi bao t r a d n g thdnh cd thfi hinh thdnh ttong giai doan sau cua qua ttinh bifit hda (sau 3 tuan cam iing, te bao bifiu hifin ca CK18, CKI9, P63 vapl-intergrin).
Difiu nay cho thdy, qua trinh chuyfin bifit hda in vitro cua T B G tmng md thanh tfi bdo bieu md CO mpt sd diem tuong ddng vdi qua trinh biet hda in vivo ttong c a thfi sdng. Vai ttd cua cdc yfiu tfi n h u KGF, E G F , H G F va IGF-2 ttong mdi tradng cdm iing cd the la diem khdi dau cho sy chuyfin bifit hda in vitro, vi ddy Id nhiing yeu to can thiet va dugc san xuat nhifiu ttong qua trinh hinh thdnh ldp bieu mo in vivo. Ngoai ra, khd ndng san xuat cdc yfiu td ty tifit vd can tifit cua tfi bdo cdm ring ciing cd thfi la mdt ly do thdc day qua trinh chuyfin biet hda xay ra bdi qua trinh bifit hda dudng n h u xay ra nhanh d cac viing cd mat dp tfi bdo cao.
KET LUAN
Te bao gdc trang md cd thfi phan lap vd dfi ddng thu nhdn tir day ron. Cdc tfi bdo nay cd the bifit hda thdnh cac tfi bao giong tfi bdo bifiu md da khi dupc nudi cdy ttong dieu kifin phii hgp, Kfit qua nghien ciru nay gop phdn cung cd thfim c a sd cho vific ting dyng lam sang vd vific tim hifiu qud trinh bifit hda ciing n h u tifim nang biet hoa ciia tfi bdo nay. T B G trang md tii day ron hi vgng se la mpt ngudn te bao ly tudng cho ky thudt md da nhan tao.
TAI LIEU THAM KHAO
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D I F F E R E N T I A T I O N O F H U M A N U M B I L I C A L C O R D - D E R I V E D M E S E N C H Y M A L S T E M C E L L S I N T O E P I D E R M A L - L I K E C E L L S
D o a n Chinh Chung'"^, Le T h a n h L o n g ^ H o a n g Nghia Son^, Nguyen H o a n g C h u o n g ' , Do M i n h Si'"', Le Van Dong^
'University of Science, VNU, HCM City
^Medical Mtiitary University, Hanoi
"'institute of Tropical Biology, VAST
Umbilical cord derived - mesenchymal stem cells (UCMSCs) can be readily obtained, avoid ethical debates or moral constraints, and show excellent plunpolency and proliferation potential. Therefore, the aim
Doan Chinh Chung etal.
of this smdy was to examine the differentiation potential of UCMSCs into epidermal cetis. In this study, UCMSCs were isolated by a tissue attachment culmre method and characterized their immunophenolype and their differentiation potency. On the third passage, UCMSCs were induced to differentiate mto epidermal cells in induction media. The results showed that UCMSCs exhibited a fibroblast-like morphology, owned strong proliferation and differentiation capacity into adipocytes and osteoblasts. Isolated UCMSCs were strong positive for CD73, CD90, CD105 and CD166, but not CD19, CD34, CD45 and HLA-DR. After differentiation, the induced cetis had morphological changes when compared with the non-induced cells, and expressed CK18, CK19, and P63 whereas non-induced cells were not. Interestingly, both induced cells and non-induced cells expressed ^1-inlergrin. These results demonsttated that UCMSCs might ttans-differentiate into epithelial cells in vilro under appropriate culmre conditions. These cells may be useful for artificial skin tissue engineering m the fumre.
Keywords: Differentiation, umbihcal cord, mesenchymal stem cells, epidermal- like cells.
Ngdy nhgn bdi: 7-3-2013
