Magnetic Field-Magnetic Nanoparticle Culture System Used to Grow In Vitro Murine Embryonic Stem Cells

Abstract

The in vitro growth of embryonic stem cells (ESCs) is usually obtained in the presence of murine embryonic fibroblasts (MEF), but new methods for in vitro expansion of ESCs should be developed due to their potential clinical use. This study aims to establish a culture system to expand and maintain ESCs in the absence of MEF by using murine embryonic stem cells (mECS) as a model of embryonic stem cell. Magnetic nanoparticles (MNPs) were used for growing mESCs in the presence of an external magnetic field, creating the magnetic field-magnetic nanoparticle (MF-MNP) culture system. The growth characteristics were evaluated showing a doubling time slightly higher for mESCs cultivated in the presence of the system than in the presence of the MEF. The undifferentiated state was characterized by RT-PCR, immunofluorescence, alkaline phosphatase activity and electron microscopy. Murine embryonic stem cells cultivated in presence of the MF-MNP culture system exhibited Oct-4 and Nanog expression and high alkaline phosphatase activity. Ultrastructural morphology showed that the MF-MNP culture system did not interfere with processes that cause structural changes in the cytoplasm or nucleus. The MF-MNP culture system provides a tool for in vitro expansion of mESCs and could contribute to studies that aim the therapeutic use of embryonic stem cells.

MgFe‐LDH Nanoparticles: A Promising Leukemia Inhibitory Factor Replacement for Self‐Renewal and Pluripotency Maintenance in Cultured Mouse Embryonic Stem Cells

“…[ 28,46 ] Magnetic nanoparticles (MNPs) can be used to maintain mESCs in the absence of MEF. [ 29 ] The intracellular depletion of iron leads to a rapid downregulation of NANOG and a dramatic decrease in the self‐renewal of human pluripotent stem cells (hPSCs), as well as spontaneous and nonspecific differentiation. [ 26 ] In this study, we proposed that the iron ion provided by MgFe‐LDH proves beneficial to the abundance of TET1/2, thus aiding the maintenance of mESC pluripotency.…”

“…Apart from the lower requirement and tolerance for aluminum ions than iron ions in mice, it has been reported that iron ions can also affect the self‐renewal of ESCs. [ 26–29 ] Hence, we proposed to replace Al ions with Fe ions for LDH modification.…”