EGF-responsive rat neural stem cells: molecular follow-up of neuron and astrocyte differentiation in vitro

J Cell Physiol. 2003 May;195(2):220-33. doi: 10.1002/jcp.10249.

Abstract

Neural stem cells (NSCs) could be very useful for the "cell therapy" treatment of neurological disorders. For this reason basic studies aiming to well characterize the biology of NSCs are of great interest. We carried out a molecular and immunocytochemical analysis of EGF-responsive NSCs obtained from rat pups. After the initial growth of NSCs as floating neurospheres in EGF-containing medium, cells were plated on poly-L-ornithine-coated dishes either in the presence or absence of EGF. We followed cell differentiation and apoptosis for 21 days in vitro and analyzed the expression levels of some genes having a major role in these processes, such as pRB, pRB2/p130, p27, and p53. We observed that EGF impairs neuronal differentiation. Furthermore, in the absence of mitogens, apoptosis, which appeared to proceed through the "p53 network," was significantly lower than in the presence of EGF. The cyclin kinase inhibitor p27, while important for cell cycle exit, seemed dispensable for cell survival and differentiation.

MeSH terms

  • Animals
  • Animals, Newborn
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Astrocytes / cytology
  • Astrocytes / drug effects
  • Astrocytes / metabolism
  • Blood Proteins / drug effects
  • Blood Proteins / genetics
  • Blood Proteins / metabolism
  • Cell Culture Techniques / methods*
  • Cell Cycle Proteins / drug effects
  • Cell Cycle Proteins / metabolism
  • Cell Differentiation / drug effects*
  • Cell Differentiation / physiology
  • Cell Lineage / drug effects*
  • Cell Lineage / physiology
  • Cell Separation / methods*
  • Cells, Cultured
  • Cyclin-Dependent Kinase Inhibitor p27
  • Epidermal Growth Factor / pharmacology*
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / physiology
  • Glial Fibrillary Acidic Protein / drug effects
  • Glial Fibrillary Acidic Protein / genetics
  • Glial Fibrillary Acidic Protein / metabolism
  • Immunohistochemistry
  • Intermediate Filament Proteins / drug effects
  • Intermediate Filament Proteins / genetics
  • Intermediate Filament Proteins / metabolism
  • Nerve Tissue Proteins*
  • Nestin
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / metabolism
  • Proteins*
  • Proto-Oncogene Proteins c-bcl-2 / drug effects
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Messenger / drug effects
  • RNA, Messenger / metabolism
  • Rats
  • Retinoblastoma Protein / drug effects
  • Retinoblastoma Protein / genetics
  • Retinoblastoma Protein / metabolism
  • Retinoblastoma-Like Protein p130
  • Stem Cell Transplantation / methods*
  • Stem Cells / cytology
  • Stem Cells / drug effects
  • Stem Cells / metabolism*
  • Tumor Suppressor Protein p53 / drug effects
  • Tumor Suppressor Protein p53 / metabolism
  • Tumor Suppressor Proteins / drug effects
  • Tumor Suppressor Proteins / metabolism

Substances

  • Blood Proteins
  • Cdkn1b protein, rat
  • Cell Cycle Proteins
  • Glial Fibrillary Acidic Protein
  • Intermediate Filament Proteins
  • Nerve Tissue Proteins
  • Nes protein, rat
  • Nestin
  • Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • Retinoblastoma Protein
  • Retinoblastoma-Like Protein p130
  • Tumor Suppressor Protein p53
  • Tumor Suppressor Proteins
  • Cyclin-Dependent Kinase Inhibitor p27
  • Epidermal Growth Factor