Hy Niloufar,

I am not sure, but I think samples of PBMC (Peripheral Blood MC) can only be obtained from circulating blood (out of a vene or artery). Bone marrow, in my opinion, would not be peripheral. The cells in bone marrow are not as much differentiated as in the blood.

I agree with Theresia. Peripheral blood mononuclear cells as the name suggests can be found only in the periphery. Cells in the bone marrow are mainly precusors that upon maturation migrate out of the bone marrow and enter the circulation. So if you want to work with PBMCs, they have to be sourced from the blood.

I wonder if you are talking strictly about isolating mononuclear cells. As explained above, PBMC is strictly in circulation and from whole blood. Mononuclear cells from blood include lymphocytes and monocytes. In the bone marrow, the isolation protocols don't differentiate between mononuclear cells and granulocytes.

Now... depending on what species you are taking these cells from pretty much determines which population will be collected.

In humans, you will almost always isolate PBMCs from whole blood using a ficoll or histopaque gradient and maybe magnetic bead purification depending on your goal.

If you are trying to isolate from mice, you will get a lot more cells from bone marrow. Most of the protocol does not generally require a gradient after flushing the femur and tibia to collect the bone marrow.

All these protocols are very standard and is easily googled.

Also, if you are differentiating the cells into macrophages or dendritic cells, the growth factors used are pretty much the same, just different concentrations.

@Niloufar

Isolation of PBMCs and Bone marrow cells is different because bone marrow contents viscosity is different as well as cellular content also vary. Protocols available from whole peripheral blood and it will not work for the same. You have to change the ratio of ficoll and blood dilutions.

Ok.thanks

have you got any protocol ?

@deepak rathore

Niloufar Mohammadkhani For human, you will need to run the BM aspirate through a mesh to exclude any clogs or large unwanted cells. From a previous user protocol, the BM aspirate are diluter in 1:3 ratio in PBS (compared to the 1:1 with peripheral blood) before using the usual density-gradient separation used for peripheral blood mononuclear cell isolation centrifuging with no brakes at RT.

For mice, bone marrow you could simply use a single-step RBC lysis (I used the one from BioLegend), which I have used before to extract T cells.

Mononuclear cells isolated from bone marrow will be less mature/differentiated than those isolated from peripheral blood. MNC isolated from bone marrow will alos contain bone marrow derived macrophages whereas PBCM isolated from peripheral blood will not contain macrophages.

@caroline

Thanks a lot dear Caroline