| Objective: Primary hepatocellular carcinoma is a difficult problem in clinic.Application of stem cell transplantation in the treatment of liver disease have beenbecoming the study focus. BMSCs (bone marrow mesenchymal stem cells, BMSCs) canbe transformed into liver stem cells and hepatocyte-like cells in the livermicro-environment. BMSCs can be chemotaxis and homing to the site of injury andpromote liver regeneration by transformated into hepatocyte-like cells under thestimulation of liver injury (such as hepatectomy). However, the role and mechanism ofBMSCs on primary hepatocellular carcinoma cells is not clear yet. In this study, aims toexplore the effect and application of BMSCs on liver cancer in microcirculation byobserving the situation of angiogenesis of hepatocellular carcinoma transplanted area.Methods: BMSCs were isolated and cultured primarily by whole bone marrowculture method, and identification of surface antigen of the3rd generation bonemarrow-derived mesenchymal stem cells by flow cytometry (FCM). Hepatoma cellscultured with BMSCs conditioned medium (BMSCs-CM) were assayed the cellproliferation rate by MTT method. The nude mice dorsal skinfold chamber liver cancermodel were established. The nude mice were divided into four groups: group A (controlgroup), group B (alone BMSCs cell transplantation group), group C (alone HepG-2cells group), group D (BMSCs and HepG-2cells co-transplanted group). Themicrovascular density of the window area at0d,7d,14d were measured by fluorescenceassay in vivo, and analyzed the results.Results: The3rd generation of bone marrow mesenchymal stem cells wasanalyzed by flow cytometry. The results showed: CD29positive rate was97.3%; CD90positive rate was98.1%; CD34positive rate was1.6%; CD45positive rate was2.9%.MTT method was used to count the proliferation rate of HepG-2cells cultured with BMSCs-CM. Through comparing with the proliferation rate of HepG-2cells culturedwith0%BMSCs-CM group,25%,50%,75%,100%BMSCs-CM group was increased(P<0.05). The nude mice dorsal skinfold chamber liver cancer model was successfullyestablished. The results of microvascular density were measured in each group at0d,7d,14d, that of group A were148.12±14.23,151.85±11.61,161.49±17.24(cm/cm2); that ofgroup B were157.01±18.94,168.98±17.99,152.93±18.17(cm/cm2); that of group Cwere165.25±25.28,173.79±29.82,211.06±40.01(cm/cm2); that of group D were151.26±16.51,183.62±22.92,248.53±29.01(cm/cm2). The result of inner groupscomparison statistical analysis showed that the microvascular density of the differenttime point were no significant difference in group A and group B and that of7dcompared with0d and14d was too in group C(P>0.05); but that of14d was higher than0d in group C(P<0.05), that of14d was higher than7d and0d in group D(P<0.05), andthat of7d was higher than0d in group D(P<0.05). The result of inter groups comparisonstatistical analysis showed that the microvascular density of the different group were nosignificant difference at0d and7d(P>0.05), but that of group C was higher than groupA and group B at14d(P<0.05), that of group D was also higher than group A and groupB at14d(P<0.05), and that of group D was also higher than group C at14d(P<0.05).Conclusion: The3rd generation BMSCs has the characteristics of stem cellsurface antigen and a strong proliferation activity. It is suitable for the study applicationin vivo and vitro. BMSCs-CM can promote the proliferation of hepatoma cells HepG-2.The hepatoma model of dorsal skinfold chamber in nude mice is establishedsuccessfully. It is observed that BMSCs can promote the growth of microvascular inhepatoma cells transplanted area using this model. |
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