원문정보
초록
영어
Embryonic stem (ES) cells are known to have an infinite proliferation and pluripotency that are associated with complex processes. The objective of this study was to examine expression of genes differentially regulated during differentiation of human ES cells by suppression subtractive hybridization (SSH). Human ES cells were induced to differentiate into neural precursor cells via embryoid body. Neural precursor cells were isolated physically based on morphological criteria. Immunocytochemical analysis showed expression of pax6 in neural precursor cells, confirming that the isolated cells were neural precursor cells. Undifferentiated human ES cells and neural precursor cells were subject to the SSH. TPX2 (Targeting Protein for Xklp2 (Xenopus centrosomal kinesin-like protein 2)) was identified, cloned and analyzed during differentiation of human ES cells into neural lineages. Expression of TPX2 was gradually down-regulated in embryoid bodies and neural precursor cells relative to undifferentiated ES cells. Targeting Protein for Xklp2 has been shown to be involved in cell division by interaction with microtubule development in cancer cells. Taken together, result of this study suggests that TPX2 may be involved in proliferation and differentiation of human ES cells.
목차
INTRODUCTION
MATERIALS AND METHODS
Culture of Human ES Cells
Differentiation of Human ES Cells into Neural Precursor Cells
Immunocytochemical Analysis
Extraction of RNA
Synthesis of cDNA by Reverse Transcription
Suppression Subtractive Hybridization
Analysis of Gene Expression by Semi-Quantitative RT-PCR
RESULTS
Identification of Differentially Expressed Genes by SSH
Un-subtracted
Down-Regulation of TPX2 Expression during Neural Differentiation
DISCUSSION
REFFRENCES