Monocyte–macrophage differentiation in three dimensional collagen lattice

Abstract

Human peripheral blood mononuclear cells (PBMC) upon transendothelial migration interact with subendothelial matrix components and differentiate into macrophages. In order to study whether the shape of the cells as dictated by the extracellular matrix can influence monocyte–macrophage (mo–mφ) differentiation, human PBMC were maintained in vitro on a three dimensional collagen I (COL I) lattice and studied for various macrophage specific functions, viz. endocytosis of [ 125I]acetyl bovine serum albumin (BSA), expression of specific cell surface antigens and expression of matrix metalloproteinases (MMPs). The cells maintained in three dimensional COL gel exhibited a higher rate of endocytosis of [ 125I]acetyl BSA than those on COL-coated plastic. FACS analysis showed that the mean fluorescence intensity (MFI) corresponding to monocyte specific LPS receptor CD14 was significantly decreased while MFI corresponding to macrophage specific transferrin receptor CD71 was significantly increased in cells maintained in vitro on three dimensional COL gel compared to two dimensional COL substrata. Expression of macrophage specific MMPs (gelatinase A and gelatinase B) was significantly high in cells maintained on COL gel than on COL I-coated plastic. Appearance of 67 kDa gelatinase in the COL gel suggested that induction as well as activation of MMPs occur when cells are maintained in a three dimensional environment. These results indicate that monocytes undergo a rapid rate of differentiation when maintained in vitro on three dimensional COL I lattice suggesting that apart from the chemical nature of the matrix, the shape of the cells as provided by the matrix also influences mo–mφ differentiation.