Fluorescent nanodiamond (FND) has been used for long-term cell labeling and in vivo cell tracking because they have good at photostability and biocompatibility. In this study, we evaluate the effect of fluorescent nanodiamond labeling on in vitro culture and differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs) into hepatocyte-like cells (HLCs). For hepatic differentiation of hUCMSCs, cells were induced with human hepatocyte growth factor, nicotinamide and Dexamethasone. FND was supplied in two experimental groups with 20 μg/mL and 100 μg/mL in 2 hours. The cell was assessed for FND uptake by laser scan microscopy and flow cytometry methods. The effect of FND on hUCMSCs was evaluated by the cell viability and growth assays as well as the differentiation throughout of morphology alterations or gene expression of anfa-fetoprotein, albumin, and hepatocyte nuclear factor 4α. The results showed that the labeling of hUCMSCs is efficient and easy and there was significant cellular uptake of FND. We did not observe any negative impacts of FND to the cell viability and growth. FND can be utilized for the long-term labeling and tracking of hUCSCs and HLCs in vivo studies. HIGHLIGHTS Fluorescent nanodiamond labeling does not affect the life and growth of hUCMSC. FND labeling does not affect the hUCMSC differentiation into hepatocyte-like cells. FND labeling maintain during the procedure of hUCMSC differentiation into hepatocyte-like cells. 2 Do, T.K.; et al.
Recent studies indicated that Mesenchymal stem cell has become a potential objective for therapy. In this study, umbilical cord cells were isolated and analyzed the expression of mesenchymal stem cells specific markers then they were differentiated into hepatocyte-like cells by DMSO and Gene transfection. Umbilical cord mesenchymal stem cell (MSC) was isolated by explant culture in media DMEM/F12, complementing with growth factors EGF, FGF and IST. After that, they were exposured to DMSO with three concentrations: 0.01%, 0.1%, 1% and another group was transfection with HNF4α by Lipofectamin LX plus. The cells were analyzed at 1, 2, 3, and 4 weeks after treatment. The cells isolation was shown the positive with markers CD73, CD34, CD86, CD90, CD105, eras, Oct 1, GATA, and negative with markers HNF4α, Alb and G6P. In group 0,1% DMSO treatment, after 3 weeks the cells were positive with markers HNF4α but it was also negative with markers Alb and G6P. In the transfection group, the cell expresses HNF4α at three weeks after treatment. Although our results exposure that the umbilical cord mesenchymal stem cells expressed hepatic specific marker after DMSO induced and DNA treatment. So it will be necessary to optimize research conditional and investigate the hepatic functions of these cells in a longer in vitro culture.
Fluorescent nanodiamond (FND) indicated that it has excellent biocompatibility and photostability, so it well suited for long-term labeling and tracking of stem cells. There are many reports concerning the factors controlling stem cell differentiation. However, still little knowledge about the biomaterials properties influence stem cell alive, growth and differentiation processing. In this study, we evaluate the effect of fluorescent nanodiamond in in vitro culture and differentiation of ucMSC into hepatocyte-like cell. Mesenchymal stem cells (MSCs) were isolated from the umbilical cord (UC) and CD markers were analyzed by flow cytometry and genes expression. For hepatic differentiation of UC-MSCs, cells were induced with HGF and DMSO treated. FND was supply in the experimental group which 10 g/ml in 4 hours. The FND uptake was detected of fluorescence intensity of FND in cells by flow cytometry and laser scan microscopy. The effect of FND into UCMSCs was not only evaluated by the cell alive and growth assay but also effective differentiation throughout morphology charging or gene expression levels of AFP, ALB, and HNF4 were determined by RT-PCR and real-time PCR. The result showed that the FND was well uptake in UCMSCs. It was no affected into ability of the cell alive and growth. The existence of FNDs does not disturb the functions of UC-MSCs differentiation into hepatocyte-like cell. FND can be utilized for the labeling and tracking of UC-MSCs and hepatocyte-like cell in homing research.
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