ABSTRACT
<p><b>OBJECTIVE</b>To induce hepatic differentiation of human adipose-derived stem cells (hADSCs) in vitro.</p><p><b>METHODS</b>hADSCs were isolated from human adipose tissue and treated with improved hepatic medium containing HGF, bFGF and FGF4. After 7 days of culture, OSM was added to the culture media. Cell growth during hepatic differentiation was evaluated by CCK8 assay. Morphology of differentiation was examined under light microscope. Liver specific genes and proteins were detected by RT-PCR analysis and immunohistochemical staining, respectively. And functional characteristics of hepatocytes were also examined.</p><p><b>RESULTS</b>The number of hADSCs cultured in the improved hepatic media was increased significantly in comparison to hADSCs cultured in control media from 5 days to 21 days (t=6.59, 8.69, 15.94 and 24.64, respectively, P<0.05). The hADSCs-derived hepatocyte-like cells exhibited hepatocyte morphology, expressed hepatocyte markers, possessed hepatocyte-specific activities, such as uptake and excretion of indocyanine green, glycogen storage and albumin production.</p><p><b>CONCLUSION</b>hADSCs can be induced into hepatocyte-like cells in this differentiation system. And this differentiation system promoted the growth of hADSCs.</p>
Subject(s)
Humans , Adipose Tissue , Cell Biology , Albumins , Metabolism , Cell Culture Techniques , Cell Differentiation , Cell Proliferation , Cell Separation , Cells, Cultured , Culture Media , Fibroblast Growth Factor 2 , Pharmacology , Hepatocyte Growth Factor , Pharmacology , Hepatocytes , Cell Biology , Metabolism , Mesenchymal Stem Cells , Cell Biology , Reverse Transcriptase Polymerase Chain Reaction , alpha-Fetoproteins , MetabolismABSTRACT
To explore a new lyophilized preservation methods for human platelets, platelets were pre-treated with aldehyde, human albumin or trehalose was added to the system of condensed cooling as protectant to stabilize the structure of platelets. The optimal resuspending buffer was also selected in the study. The morphological changes of platelets were observed by using electron microscopy after lyophilization, and the expression of membrane proteins on platelets was detected also after lyophilization. The results indicated that the recovery rate of platelets treated with aldehyde was generally more than 60%. Aggregative ability was reduced a little than the platelet untreated. 5% of human albumin had an advantage over 40 mmol/L of trehalose in respect of the preservation effect. In the way of keeping aggregative ability, PPP was obviously better than PBS. The results of electron microscopy displayed that organelles including mitochondria and excreted granules could be observed distinctly. Whereas, expression of membrane proteins of platelet treated with aldehyde was evidently dropped as compared with those of the fresh platelet. In conclusion, aldehyde as a novel protective agent, has excellent effects on lyophilization of platelets and is worthy to be further studied.
Subject(s)
Humans , Albumins , Pharmacology , Aldehydes , Pharmacology , Blood Platelets , Cell Biology , Blood Preservation , Methods , Freeze Drying , Reproducibility of Results , Trehalose , PharmacologyABSTRACT
Platelet is activated through signal transduction, that mainly includes phospholipase-beta (PLCbeta) pathway, protein tyrosine kinases (PTK) pathway, phosphatidylinositol3-kinase (PI3-K) pathway, mitogen-activated protein kinases (MAPK) pathway, cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) pathway and phospholipase A2 (PLA2) pathway. This article focuses on the relationship between signal transduction and platelet activation.