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Updated: Mar 9, 2026
Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis
Published on: December 18, 2019
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.
Background:
Chordoma are malignant tumors of the axial skeleton, which arise from remnants of the notochord. The Notochord (chorda dorsalis) is an essential embryonic structure involved in the development of the nervous system and axial skeleton. Therefore, the notochord seems to be the most biologically relevant control tissue to study chordoma in molecular biology research. Nevertheless, up to now mainly different tissues but not the notochord have been used as control for chordoma, due to difficulty of isolating notochordal tissue. Here, we describe a fast and precise method of isolating notochordal cells.
Methods:
Examination of human fetuses, with a gestation of 9, 11 and 13 weeks, using (immuno)histochemical methods was performed. To isolate pure notochord cells for further molecular biology investigation five flash frozen fetuses between 9 and 10 weeks of gestation were dissected by microtome slicing. Thereafter pure notochord cells for further molecular biology investigation where harvested by using laser capture microdissection (LCM). RNA was extracted from these samples and used in quantitative PCR.
Results:
This study illustrates notochord of embryonic spines in three different stages of gestation (9-11-13 weeks). Immunohistochemical staining with brachyury showed strong staining of the notochord, but also weak staining of the intervertebral disc and vertebral body. LCM of notochord slices and subsequent total RNA extraction resulted in a good yield of total RNA. qPCR analysis of two housekeeping genes confirmed the quality of the RNA.
Conclusion:
LCM is a fast and precise method to isolate notochord and the quality and yield RNA extracted from this tissue is sufficient for qPCR analysis. Therefore early embryo notochord isolated by LCM is suggested to be the gold standard for future research in chordoma development, classification and diagnosis.
Insights
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.
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.
