EFFECTS OF MESENCHYMAL STEM CELL TRANSPLANTATION INTO ISCHEMIC BILE DUCT INJURY TISSUES OF RATS ON THE EXPRESSION OF VASCULAR ENDOTHELIAL CELL PHENOTYPE AND THE INCREASE OF VASCULAR DENSITY
DOI:
https://doi.org/10.26873/SVR-844-2019Abstract
We aimed to evaluate the effects of mesenchymal stem cell (MSC) transplantation into the ischemic bile duct injury tissues of rats on the expression of vascular endothelial cell phenotype and the increase of vascular density. The bone marrow of male Wistar rats aged 2-4 weeks was sampled to culture MSCs by the direct adhesion method. Forty-six female rats were randomly divided into a transplantation group (group A) and a non-transplantation group (group B). Group A was transplanted with MSCs (1×106 cells/site, a total of 8 sites) into ischemic biliary lesion. After 21 days, the bile ducts with lesions were removed. The transplanted MSCs were tracked by a combination of in situ hybridization staining of sex-determining region Y gene with CD34 immunohistochemical staining (double labeling). Microvascular and vascular endothelial growth factors were labeled by immunohistochemical staining to calculate microvascular density (MVD). After 21 days of injection, double labeling revealed that a small portion of Y-chromosome positive cells in group A (brown cell nucleus) had CD34 antigen phenotype (bluish violet). MVD of group A ((63 ± 18)/HP) was higher than that of group B ((53 ± 14)/HP), and the vascular endothelial growth factor (VEGF) expressions of the two groups were significantly different (P<0.05). The formation of vascular endothelial cells after MSC transplantation improved the blood supply of bile duct by secreting VEGF.
Key words: mesenchymal stem cell; transplantation; in situ hybridization; immunohistochemistry; bile duct injury
UČINKI TRANSPLANTACIJE MEZENHIMALNIH MATIČNIH CELIC V ISHEMIJSKO TKIVNO POŠKODBO ŽOLČNIH KANALOV NA IZRAŽANJE ŽILNEGA ENDOTELIJSKEGA FENOTIPA CELIC IN POVEČANJE CELIČNE GOSTOTE
Povzetek: V študiji smo želeli oceniti učinke presaditve mezenhimskih matičnih celic (MSC) v ishemična tkiva poškodovanih žolčnih kanalov pri podganah na izražanje fenotipa žilnih endotelijskih celic in na žilno gostoto. Celice kostnega mozga podganjih samcev seva Wistar, starih 2 – 4 tedne smo gojili v celični kulturi v ustreznih gojiščih. Šestinštirideset samic podgan je bilo naključno razdeljenih v presaditveno skupino (skupina A) in ne-presaditveno skupino (skupina B). Skupini A smo presadili MSC (1 × 106 celic/mesto, skupaj 8 mest) v ishemično poškodbo žolčnih kanalov. Po 21 dneh so bili poškodovani žolčni kanali odvzeti za histološke preiskave. Presajene MSC so bile spremljane s kombinacijo dveh metod - hibridizacije in situ, s katero smo označili gen Y za določanje spola, ter z imunohistokemičnim barvanjem proti beljakvoini CD34 (dvojno označevanje). Mikrovaskularni in vaskularni endotelni rastni faktorji so bili označeni z imunohistokemičnim barvanjem, na podlagi katerega smo izračunali mikrovaskularno gostoto (MVD). Enaindvajset dni po injiciranju je dvojno označevanje pokazalo, da je majhen del celic s kromosomov Y (presajenih celic) v skupini A izražal antigen CD34. MVD skupine A (63 ± 18)/HP) je bil večji od MVD v skupini B ((53 ± 14)/HP) in izražanje žilnih endotelnih rastnih faktorjev (VEGF) se je statistično značilno razlikovalo med skupinama (P < 0,05). Tvorba žilnih endotelnih celic po presaditvi MSC je izboljšala preskrbo žolčnih kanalov s krvjo preko izločanja VEGF.
Ključne besede: mezenhimske matične celice; presaditev; hibridizacija in situ; imunohistokemija; poškodba žolčnih kanalov
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