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Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis
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Notochord isolation using laser capture microdissection.

R G C Santegoeds1, Y Yakkioui1, A Jahanshahi2

  • 1Department of Neurosurgery, Maastricht University Medical Center, Maastricht, The Netherlands; Department of Neuroscience, Maastricht University Medical Center, Maastricht, The Netherlands.

Journal of Chemical Neuroanatomy
|December 29, 2016
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method using laser capture microdissection (LCM) to isolate notochord cells. This technique provides high-quality RNA for studying chordoma, a rare bone cancer, and is recommended as a gold standard.

Keywords:
ChordomaHuman embryoLaser-capture microdissectionNotochordTissue dissection

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Area of Science:

  • Oncology
  • Developmental Biology
  • Molecular Biology

Background:

  • Chordoma is a rare malignant tumor originating from notochord remnants in the axial skeleton.
  • The notochord is crucial for embryonic development of the nervous system and axial skeleton.
  • Studying chordoma requires the notochord as a control tissue, but its isolation is challenging.

Purpose of the Study:

  • To establish a precise and efficient method for isolating notochordal cells from embryonic tissue.
  • To validate the suitability of isolated notochordal cells for molecular biology research, particularly for chordoma studies.

Main Methods:

  • Human fetuses (9-13 weeks gestation) were examined using immunohistochemical methods.
  • Microtome slicing and laser capture microdissection (LCM) were employed to isolate pure notochordal cells.
  • Total RNA was extracted from isolated cells for quantitative PCR (qPCR) analysis.

Main Results:

  • Immunohistochemical staining confirmed notochordal presence and identified brachyury expression.
  • LCM successfully isolated notochordal cells, yielding sufficient high-quality RNA.
  • qPCR analysis confirmed the integrity and quality of the extracted RNA.

Conclusions:

  • Laser capture microdissection (LCM) is a rapid and accurate method for isolating embryonic notochord.
  • The RNA yield and quality from LCM-isolated notochord are adequate for qPCR.
  • Embryonic notochord isolated by LCM should be considered the gold standard for future chordoma research.