Abstract
Mesenchymal stem cells (MSCs) have various functions, making a significant contribution to tissue repair. On the other hand, the viability and function of MSCs are not lasting after an in vivo transplant, and the therapeutic effects of MSCs are limited. Although various chemical modification methods have been applied to MSCs to improve their viability and function, most of conventional drug modification methods are short-term and unstable and cause cytotoxicity. In this study, we developed a method for long-term drug modification to C3H10T1/2 cells, murine mesenchymal stem cells, without any damage, using the avidin-biotin complex method (ABC method). The modification of NanoLuc luciferase (Nluc), a reporter protein, to C3H10T1/2 cells by the ABC method lasted for at least 14 days in vitro without major effects on the cellular characteristics (cell viability, cell proliferation, migration ability, and differentiation ability). Moreover, in vivo, the surface Nluc modification to C3H10T1/2 cells by the ABC method lasted for at least 7 days. Therefore, these results indicate that the ABC method may be useful for long-term surface modification of drugs and for effective MSC-based therapy.
Highlights
Mesenchymal stem cells (MSCs) are generally isolated from a variety of tissues including bone marrow, adipose tissue, umbilical cord blood, and placenta[1,2,3]
Genetic engineering methods are frequently applied to various cells and the engineered MSCs may be effective in the treatment of various diseases[16,17,18], some disadvantages remain: 1) low transfection efficacy, 2) lengthy cultivation for the establishment of a stable gene-expressing clone, and 3) risks associated with viral vectors
The strong fluorescence was observed only on the surface of fluorescent streptavidin-modified C3H10T1/2 cells and green fluorescent protein (GFP)-modified C3H10T1/2 cells, whereas slight signals were observed in C3H10T1/2 cells
Summary
Mesenchymal stem cells (MSCs) are generally isolated from a variety of tissues including bone marrow, adipose tissue, umbilical cord blood, and placenta[1,2,3]. The function and viability of MSCs are generally transient and low after an in vivo transplant This is because most of transplanted MSCs may disappear under the influence of immune cells and via negative effects of endogenous or environmental changes (inflammation, ischemia-reperfusion, and the lack of nutrition and oxygen)[12,13]. To overcome these disadvantages of MSC transplantation, recent studies have shown that genetic engineering or surface chemical modification improves and diversifies the therapeutic potential of MSCs12,14,15. The in vivo duration of surface modification of C3H10T1/2 cells was evaluated in nude mice by means of an in vivo imaging system
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