MacroH2A1 Regulation During the Cell Cycle of Mouse Embryonic Stem Cells

MacroH2A is a core histone variant that plays an important role in the X-inactivation process during differentiation of embryonic stem cells. It has been shown that macroH2A changes in localization during the cell cycle of somatic cells. This study aims to determine how macroH2A changes during the cell cycle of embryonic stem cells. Male and female mouse embryonic stem cells were transfected with a GFP::macroH2A construct and the relationship between macroH2A and the cell cycle was determined using FACS. This study shows that macroH2A is altered during the cell cycle of embryonic stem cells as it is in somatic cells and that in randomly cycling cells, there is a correlation between macroH2A expression and the phases of the cell cycle. High GFP expressing cells are mostly in the G2/M phase and low GFP expressing cells are mostly in the G1 phase. This correlation indicated that macroH2A is replicated with cellular DNA during the S phase resulting in higher expression in the G2/M phase. Future research, such as RT-PCR and differentiation experiments, is needed to further study this relationship and determine whether this change is at the protein or RNA level and how it changes during differentiation.

Determination of the Myogenic Potential of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells

Human embryonic stem cells (hESCs) have the potential to differentiate to all adult somatic cells. This property makes hESCs a very promising area of research for the treatment of disorders in which specific cell populations need to be restored. Despite this potential, research that focuses on producing mesodermally derived cell populations from hESCs is decidedly limited, notwithstanding the prevalence of disorders involving mesodermal tissues for which treatment options are limited. Skeletal muscle myoblasts are derivatives of mesodermal cells and are characterized by the expression of the MyoD gene. These cells are difficult to obtain from hESCs in a reproducible and efficient manner. Recent developments in the field have showed some success in obtaining myogenic cells from hESCs through a mesenchymal stem cell (MSC)-like intermediate population. MSCs, which are an adult stem cell population typically derived from the bone marrow, are capable of generating multiple cell types including skeletal muscle. The aim of this study was to develop an efficient method that derives myoblasts from an MSC-like intermediate. To accomplish this goal, we first set out to isolate and expand the MSC-like intermediate from hESCs differentiated in vitro. Difficulties in reproducing published cell-differentiation methodologies, which represent a significant and familiar challenge in hESC research, are highlighted in this report.