ABSTRACT
Human parthenogenetic embryonic stem cells (hpESCs) established from artificially activated oocytes have a wider immune-matching ability because of their homozygosity in the major histocompatibility complex alleles. Whether these cells possess the differentiation capacity similar to regular human embryonic stem cells (hESCs) derived from fertilized eggs is unclear. The aims of this study were to determine whether hpESCs could be differentiated into multipotent mesenchymal stem cell (MSC)-like cells in vitro and then compare these cells with those derived from hESCs. MSC-like cells were obtained from both hpESCs and hESCs, which exhibited similar cell surface marker expression profiles. Further analyses revealed that cells derived from hpESCs possessed stronger osteogenic but weaker adipogenic potentials compared with cells derived from hESCs. This is the first work that demonstrates the differentiation of hpESCs into multipotent MSC-like cells. These hpESCs could be a potential source for cell-based therapies.
Subject(s)
Cell Differentiation , Embryonic Stem Cells/physiology , Mesenchymal Stem Cells/physiology , Alkaline Phosphatase/metabolism , Antigens, Differentiation/metabolism , Cells, Cultured , Coculture Techniques , Collagen Type II/metabolism , Embryoid Bodies , Embryonic Stem Cells/metabolism , Fibroblasts/cytology , Humans , Intermediate Filament Proteins/metabolism , Lipid Metabolism , Mesenchymal Stem Cells/metabolism , Muscle Proteins/metabolism , Nerve Tissue Proteins/metabolism , Nestin , Vimentin/metabolismABSTRACT
Human embryonic stem (ES) cells have been established either from fresh or frozen embryos. The recovery rates of undifferentiated human ES cells after cryopreservation with conventional slow-rate freezing and rapid-thawing methods are relatively low. The purpose of this study was to improve cryopreservation efficiency by modifying conventional methods with addition of trehalose. Immature oocytes donated from patients undergoing IVF treatment were utilized to generate blastocysts. One human ES cell line (named hES1) was established and characterized in detail. The hES1 cells expressed regular human ES cell markers, including stage-specific embryonic antigens SSEA-3, SSEA-4, tumour rejection antigens TRA-1-60, TRA-1-81 and octamer-binding transcription factor Oct-4 with high levels of alkaline phosphatase and telomerase activities. Cells could be differentiated to form teratomas in vivo. With slow-rate freezing and rapid-thawing methods modified by adding trehalose, the recovery rate of undifferentiated hES1 cells has been greatly improved from 15 to 48%. Cells retained pluripotency with normal karyotype after thawing. The results indicated that the use of trehalose is efficient and convenient for cryopreservation of human ES cells.
