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Differential Expression of TPX2 upon Differentiation of Human Embryonic Stem Cells

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

ABSTRACT
 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

저자정보

  • Hye Min Noh Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University
  • Seong-Jun Choi Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University
  • Sehee Kim Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University
  • Kye-Seong Kim College of Medicine, Hanyang University
  • Jin Kyeoung Kim Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University

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