Gene Expression Profiling in Laser-Microdissected Bone Marrow Megakaryocytes
Gene expression analysis of the megakaryocytic lineage requires isolation of megakaryocytes from their bone marrow microenvironment. Laser microdissection of megakaryocytes from diagnostic bone marrow biopsies allows analysis of standardised formalin-fixed samples that reflect the in situ grown status quo of a physiological or pathological condition. Taking into account that in neoplastic proliferation, e.g. myeloproliferative neoplasms, non-neoplastic haematopoietic clones proliferate in parallel, this morphology-based isolation enables selective analysis of the aberrant megakaryocytic population. Two different laser microdissection devices are presented, and the details of RNA extraction and sub-sequent real-time qPCR gene expression analysis of mRNA and microRNA are provided.
Key wordsLaser microdissection Bone marrow Megakaryocyte Gene expression Real-time qPCR RNA microRNA
The author would like to thank his mentors Professor Dr. med. Hans H. Kreipe and Professor Dr. med. Oliver Bock (Institute of Pathology, Hannover Medical School) as well as Dr. phil. Khadra Hussein, Dr. med. Ulrich Thorns and Sir Jan Off.
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