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Updated: May 23, 2026
Culturing Microglia from the Neonatal and Adult Central Nervous System
Published on: August 9, 2013
Microglia differentiation using a culture system for the expansion of mice non-adherent bone marrow stem cells
Arnd Hinze1, Alexandra Stolzing
1Fraunhofer Institute for Cell Therapy and Immunology, Perlickstraße 1, 04103, Leipzig, Germany. alexandra.stolzing@izi.fraunhofer.de.
Introduction:
Studying primary adult microglia is hampered because of the difficult isolation procedure and the low cell yield. We therefore established a differentiation protocol using a culture system developed for the expansion of non-adherent bone marrow cells.
Methods:
Non-adherent bone marrow derived stem cells (NA-BMC) are derived by selective adhesion ('preplating') and are non adhesive adult stem cells. We investigated the changes in bone marrow cell populations by this repeated selective adhesion and compared the potential of the derived cells to differentiate towards microglia. Cells were differentiated with astrocyte conditioned medium (ACM) and granulocyte-monocyte colony stimulating factor (GM-CSF).
Results:
NA-BMC cultures show a steep raise in the fraction of stem cells during the cultivation time and the differentiation potential is of the same quality as established protocols. Around 70% of the cells are microglia defined as being positive for CD11b/CD45 and show phagocytosis activity and oxidative bursts.
Conclusion:
The non-adherent cell system has the advantage that is produces stem cell progenitors during expansion and provides good microglial differentiation.
Insights
This study presents a new method for isolating adult microglia using non-adherent bone marrow stem cells. This approach yields a high percentage of microglia with key functional characteristics.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Immunology
Background:
- Primary adult microglia isolation is challenging due to difficult procedures and low cell yields.
- A novel differentiation protocol was developed using a culture system for non-adherent bone marrow cells.
Purpose of the Study:
- To establish an efficient method for obtaining adult microglia.
- To compare the microglial differentiation potential of cells derived from a non-adherent bone marrow stem cell system.
Main Methods:
- Non-adherent bone marrow derived stem cells (NA-BMC) were isolated via selective adhesion (preplating).
- Cells were differentiated into microglia using astrocyte conditioned medium (ACM) and granulocyte-monocyte colony stimulating factor (GM-CSF).
- Changes in bone marrow cell populations and microglial differentiation potential were investigated.
Main Results:
- NA-BMC cultures demonstrated an increased fraction of stem cells during cultivation.
- Approximately 70% of differentiated cells were identified as microglia (CD11b/CD45 positive).
- These microglia exhibited characteristic phagocytosis activity and oxidative bursts.
Conclusions:
- The non-adherent cell system facilitates the expansion of stem cell progenitors.
- This method provides a robust approach for microglial differentiation, overcoming previous isolation challenges.
