Efficient Isolation and Long-term Red Fluorescent Nanodia-mond Labeling of Umbilical Cord Mesenchymal Stem Cells for the Effective Differentiation into Hepatocyte-like Cells

Abstract

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.

Keywords:
Differentiation; fluorescent nanodiamond; hepatocyte-like cells; in vitro culture; umbilical cord stem cells