Effects of extracellular matrix proteins on macrophage differentiation, growth, and function: comparison of liquid and agar culture systems.
Both spaceflight and skeletal unloading suppress the haematopoietic differentiation of macrophages (Sonnenfeld et al., Aviat. Space Environ. Med., 61:648-653, 1990; Armstrong et al., J. Appl. Physiol., 75:2734-2739, 1993). The mechanism behind this reduction in haematopoiesis has yet to be elucidated. However, changes in bone marrow extracellular matrix (ECM) may be involved. To further understand the role of ECM products in macrophage differentiation, we have performed experiments evaluating the effects of fibronectin, laminin, collagen type I, and collagen type IV on macrophage development and function. Bone marrow-derived macrophages cultured on four different ECM substrates in liquid culture medium showed less growth than those cultured on plastic. Significant morphological differences were seen on each of the substrates used. Phenotypically and functionally, as measured by class II major histocompatibility molecule (MHCII) expression, MAC-2 expression, and the secretion of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha), these macrophages were similar. In contrast, bone marrow-derived macrophages cultured in suspension, using agar, showed no difference in growth when exposed to ECM proteins. However, IL-6 and TNF-alpha secretion was affected by fibronectin, laminin, collagen type I, and collagen type IV in a concentration-dependent manner. We conclude that the ECM products fibronectin, laminin, collagen type I, and collagen type IV have profound effects on macrophage development and function. Additionally, we suggest that an ECM-supplemented agar culture system provides an environment more analogous to in vivo bone marrow than does a traditional liquid culture system.
Division of Biology, NASA Specialized Center of Research and Training, Kansas State University, Manhattan 66506.
J W
JW
Armstrong
S K
SK
Chapes
D000362:Agar; D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis; D001854:Bone Marrow Cells; D002455:Cell Division / Q000502:physiology; D003470:Culture Media; D016326:Extracellular Matrix Proteins / Q000502:physiology*; D005260:Female; D037502:Galectin 3; D006410:Hematopoiesis / Q000502:physiology; D000949:Histocompatibility Antigens Class II / Q000096:biosynthesis; D006801:Humans; D016130:Immunophenotyping; D015850:Interleukin-6 / Q000378:metabolism; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000943:Antigens, Differentiation; D003470:Culture Media; D016326:Extracellular Matrix Proteins; D037502:Galectin 3; D000949:Histocompatibility Antigens Class II; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D000362:Agar