Isolation of murine bone marrow by centrifugation or flushing for the analysis of hematopoietic cells – a comparative study

Abstract

Investigation of the bone marrow as the main compartment of hematopoiesis is critical in many research fields. Here, we adapted a centrifugation-based method for the isolation of murine bone marrow and compared it to the traditional flushing method. Analysis of primary hematopoietic stem cells, immune cells, and megakaryocytes revealed a comparable distribution of cellular (sub)populations. Furthermore, in vitro differentiated megakaryocytes displayed unaltered proplatelet formation. Strikingly, bone marrow isolation by centrifugation was considerably faster than the flushing method and significantly increased the cell yield. Thus, the centrifugation-based isolation method is highly suitable for the study of murine bone marrow cells.

Keywords:

• Bone marrow
• hematopoietic stem cells
• immune cells
• megakaryocytes
• proplatelet formation

Acknowledgements

We thank Lou M. Wackerbarth for help with megakaryocyte cultivation experiments.

Authorship Contribution

TH and CG performed experiments, analyzed data, and wrote the manuscript; MLM established the immune cell flow cytometry in our laboratories; IP and DS designed and supervised research, and wrote the manuscript.

Declaration Of Interest

DS is on the editorial board of Platelets.

Additional information

Funding

This study was supported by the Deutsche Forschungsgemeinschaft (SFB/TR 240, project number 374031971, project B08 to DS and A01 to IP) and the European Union (Thrombo-Inflame, EFRE–Europäischer Fonds für regionale Entwicklung, Bavaria).