Stably transfected human embryonic stem cell clones express OCT4-specific green fluorescent protein and maintain self-renewal and pluripotency

摘要: Human embryonic stem cells (hESCs) are derived from the inner cell mass of preimplantation embryos; they can be cultured indefinitely and differentiated into many types in vitro. These therefore have ability to provide insights human disease a potential unlimited supply for cell-based therapy. Little is known about factors that important maintaining undifferentiated hESCs vitro, however. As tool investigate these factors, transfected hES clonal lines were generated; able express enhanced green fluorescent protein (EGFP) reporter gene under control OCT4 promoter. an marker state central regulator pluripotency ES cells. OCT4-EGFP exhibit features similar parental hESCs, pluripotent, produce all three germ layer Expression colocalized with endogenous OCT4, as well hESC surface antigens SSEA4 Tra-1-60. In addition, expression retained culture extensive period time. Differentiation toward neural lineage targeted knockdown by RNA interference downregulated EGFP lines, this correlates closely reduction expression. Therefore, easy noninvasive method monitor will invaluable studying not only function self-renewal differentiation but also required maintenance culture.