Long-Term Bone Marrow Culture

Abstract

Dugotrajna kultura koštane srži eksperimentalni je model hematopoeze koji in vitro reproducira uvjete hematopoetskog mikrookružja in vivo. U dugotrajnoj kulturi postiže se proliferacija i diferencijacija primitivnih matičnih hematopoetskih stanica mezenhimskog podrijetla. Dugotrajnu kulturu koštane srži čine adherentni stromalni sloj i slobodne stanice u tekućem dijelu (supernatantu) kulture. U dugotrajnoj se kulturi hematopoeza zbiva bez egzogenih poticatelja rasta jer njih stvara stromalni sloj. Izgled i razvitak strome, proliferacija i diferencijacija progenitorskih stanica i proizvodnja citokina razlikuju se u dugotrajnoj kulturi normalne i patološki promijenjene koštane srži. Dugotrajna se kultura koštane srži upotrebljava u fundamentalnim istraživanjima hematopoeze, u farmakološkim ispitivanjima učinaka lijekova i imunocitokina na hematopoezu, za čišćenje autotransplantata koštane srži od preostalih leukemijskih stanica te za prijenos gena. Također je prikladan laboratorijski model za in vitro ispitivanje karcinogenih i drugih štetnih činitelja iz okoline koji utječu na krvotvorno tkivo pri profesionalnom ili habitualnom izlaganju.Long-term bone marrow culture is an experimental in vitro model of hematopoiesis imitating conditions in vivo. It contains hematopoietic elements at various stages of differentiation as well as a supportive stromal microenvironment. Primitive hematopoietic stem cells of mesenchymal origin, the long-term culture initiating cells proliferate and differentiate into different cell types, giving rise to the adherent stromal layer and to various hematopoietic elements attached to it or floating freely in the supernatant medium. The stromal layer keeps the hematopoietic cells aggregated, helps their mitosis, differentiation and maturation by cell-to-cell contact, produces hematopoietic growth factors (cytokines), and forms the extracellular matrix required for cell attachment. Hematopoiesis occurs without exogenous growth factors. The appearance and development of the stroma, the proliferation and differentiation of hematopoietic progenitor cells, and the production of cytokines differ in long-term cultures of the normal and of pathologically altered bone marrow. Long-term bone marrow culture is applied in fundamental studies of normal and pathologically altered hematopoiesis, in pharmacological research, in the purging of residual leukemia cells from bone marrow autotransplants, and in the gene transfer. It is also suitable for testing carcinogenic and toxic chemicals causing hematopoietic damage through occupational or habitual exposure