Volume 60 (2010) Issue: 2010 No#1

Cultivation of hamster bone marrow haematopoietic stem and progenitor cells

Author(s): Kovačević Filipović Milica, Okić Ivana, Petrićević Tanja, Mojsilović S, Krstić Aleksandra, Jovčić Gordana, Bugarski Diana, Milenković P, Petakov Marijana, Radovanović Anita, Božić Tatjana, Ivanović Z

Keywords:hamster, hematopoiesis, progenitor cell, stem cell, commercial methylcellulose media

Hamster, a hibernating animal, is an important experimental model in research on the influence of hypothermia on different physiological processes. A simple procedure for cultivation and identification of hamster hematopoetic stem cells (HSC) and hematopoetic progenitor cells (HPC) is a premise for a succesfull investigation upon hypothermia effects on hematopoiesis. The aim of this work was to evaluate the utilization of commercially available methylcellulose media (MC) and recombinant mouse and human cytokines for hamster HSC and HPC assays, in order to enable further studies on these cells. Hamster bone marrow mononuclear cells (BMMNC) were plated in MC containing cytokines that support mouse or human HPC growth. Also, BMMNC were resuspended in cytokine supplemented liquid media and incubated for 5 weeks with a four day monitoring of viable cell number. We demonstrated that hamster hematopoietic progenitor cells committed for erythroid lineage and myeloid lineage successfully formed recognizable colonies in both mouse and human MC, while multipotent progenitor cells formed colonies only in mouse MC. We also defined conditions for the evaluation of hamster HSC activity in liquid cultures, based on continuous 5 weeks HSC proliferation. The obtained results verify the utilization of mouse specific MC for further research on hamster HPC biology during hypothermia.