Effects of a preformed extracellular matrix on long-term serum-free bone marrow culture.
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.
Teofili
L
L
Istituto di Semeiotica Medica, Università Cattolica del Sacro Cuore, Rome, Italy.
Sargiacomo
M
M
Iovino
M S
MS
Zini
G
G
Leone
G
G
Bizzi
B
B
Peschle
C
C
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
D003470:Culture Media