title: Effects of a preformed extracellular matrix on long-term serum-free bone marrow culture.
abstract: The extracellular matrix (ECM) produced by the stromal layer plays a key role in the regulation of commitment and differentiation of hematopoietic cells. Long-term bone marrow culture (LTBMC) allows analysis of the stromal microenvironment. Recently, serum-free LTBMC has been described, but the formation of a classical adherent layer was never observed under these conditions. We have evaluated the effect(s) of a chemically well defined ECM on serum-free and serum-dependent LTBMC. In serum-dependent cultures ECM did not induce a significant increase of hematopoiesis. In serum-free conditions, a marked improvement of hematopoiesis was observed, both in terms of CFU-GM and BFU-E yield and in duration of cultures. A confluent stromal layer was observed only in the presence of ECM. The present results indicate that the addition of ECM to serum-free cultures provides a standardized culture condition, while improving progenitor cell recovery and allowing formation of a confluent stromal layer. Moreover, ECM+LTBMC may provide a model to study the effect(s) of adhesive proteins and hematopoietic growth factors normally present in serum.
authors.affiliation: Istituto di Semeiotica Medica, Università Cattolica del Sacro Cuore, Rome, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Teofili
authors.forename: M
authors.initials: M
authors.lastname: Sargiacomo
authors.forename: M S
authors.initials: MS
authors.lastname: Iovino
authors.forename: G
authors.initials: G
authors.lastname: Zini
authors.forename: G
authors.initials: G
authors.lastname: Leone
authors.forename: B
authors.initials: B
authors.lastname: Bizzi
authors.forename: C
authors.initials: C
authors.lastname: Peschle
mesh_terms: D001769:Blood; D001853:Bone Marrow / Q000502:physiology*; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D003470:Culture Media; D005109:Extracellular Matrix / Q000502:physiology*; D006801:Humans; D013997:Time Factors
chemical_list: D003470:Culture Media