Dear All,
I am deciding between two methods to isolate BMDM and I need input please of which could be better.
Method 1: Isolating BM macrophage precursors.
Chapter Murine Bone Marrow-Derived Macrophages
This method is based on isolating macrophage bone marrow precursors. The advantage is that differentiating macrophages from precursors yield more homogeneous cultures.
Method 2: Isolating and plating BM cells directly.
The other one is the standard procedure (attached in file) that involves isolating BM cells and plating them for 7 days in MCSF-1.
The advantage of this methods is that it is shorter and uses less resources.
Which one would you use?
thank you,
Matias.
I guess that depends on your experimental context. I am not sure how the precursor cells are defined. Regardless, the frequencies of precursor cells in the bone marrow are super low and it may take a lot of precursor cells/ time and effort to get sufficient yield. Also isolating precursors might need specialized equipment and it is labor intensive. The widely used strategy is to plate BM cells with M-CSF. This (like you said) would lead to a heterogeneous mix of macrophages, which are derived from monocytes and dendritic cells. In summary, you need to choose the method based on what is important to you- a pure homogeneous population or higher yield but a less homogeneous population. Keep in mind that using whole BM might be more physiologically relevant than isolating precursors and differentiating them to macrophages.
I agree with Julie Joseph in that it depends on your experimental context. If you're curious about macrophages derived from a particular precursor, then you're going to need to sort that precursor cell. If you're just looking for a general macrophage culture, then I would just do the BMDM method. It's relatively easier and generates lots of cells to work with.
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