We want to obtain red bone marrow for the assessment of iron stores. Bone marrow is obtained using a biopsy needle during orthopedic knee and hip surgeries, in which the femur is accessible. However, we often end up with white bone marrow, including high amounts of fat and useless for the assessment of iron stores. Does anyone know the a good (or the best) method to solely obtain red bone marrow? Unfortunately, we can not change or biopsy site.
If you cannot change the biopsy site then you have to change the age of your probants. Only the child's distal femur still contains red bone marrow. In adults it is contained in flat bones(sternum, etc)and event
The more distal you biopsy the femur and the older the individual the less red bone marrow you will aspirate. I agree with Panayot Tanchev.
Do you have a possibility to aspirate a larger volume?
Consider the possibility to enrich red marrow cells by centrifugation or sorting. However, such strategy requires information about your specific question.
The age of patient is a real problem (see attached file). I think that you use for the assessment of iron stores the Perls stain on the bone marrow frustules. I suggest you two way to obtain a god material: i) you can transfer the bone marrow into a Petri plate, search visually and take the single frustules and, finally you can crash or smear those on the slide; ii) you can performe 2 consecutive wash cycles with PBS in order to eliminate interference by microfibers and fat droplets.
Dear all, thank you very much for the answers and helpfull suggestions! We are bounded by age and biopsy site (must be easily accessible during total hip or knee replacement). Indeed, we assess iron stores using the Perls stain on the bone marrow frustules, and already use a Petri plate to selects the frustules. However, they are hard to find and we often do not find any of them at all. We aspirate 4mL of bone marrow (+ 1 mL of EDTA), but we can, and will, increase the amount. Also we will try your suggestion, Antonio La Gioia, and try to obtain frustules after 2 consecutive wash cycles with PBS. I do not have any experience with this, however, I think the laboratory will. Are there any other ways to select the frustules for the Perls Prussian Blue staining? Also, is the change of obtaining red bone marrow higher at the proximal femur (during hip replacement) compared to the distal femur? Again, thank you all very much for the help!
Proximal femur still contains red marrow also in adults. You may obtain a lot of marrow from resected head and neck after THR. One obstacle may be if the surgeons send routinely the resected head to the bone bank ?
I do not know if they send it to the bone bank. However, is it still possible to obtain the frustules from the resected head and neck? I can imagine the bone marrow inside will quickly coagulate and selecting and extracting the iron containing particles might get impossible.
I think this is a problem of good collaboration with the surgeons. You may get the femoral head as soon as it is resected.
Dear Niels
4 mL of bone marrow is a very high amount but if you can still increase the aspiration, consider the possibility to enrich red marrow cells by centrifugation, as suggested by Dr Jirko, before washing and/or transfer into Petri plate. In addition you can pre-select the patients by performing serum ferritin and descarding those whit low ferritin's levels.
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