Role of the extracellular matrix on the growth and differentiated phenotype of murine colonic adenocarcinoma cells in vitro

Abstract

The growth and differentiation characteristics of MAC 15 murine adenocarcinoma cells, derived from routine passage in vivo for growth in vitro on a plastic substrate (MAC15j cells), were compared under conditions in which the cells were seeded onto a substrate of type-I collagen which was either attached to plastic or was released to float free in medium. Cells grown on a plastic substrate consisted of a heterogeneous, largely anaplastic population with a putative enterocytic morphology but with no evidence of junctional complexes or cell polarity typical of an epithelial phenotype. MAC 15j cells from cultures grown on a plastic substrate reestablished a moderate to well-defined degree of differentiation when transplanted back into NMRI mice. When MAC 15j cells were seeded from plastic onto type-I collagen, either attached to plastic or free-floating, tight junctional complexes were formed and the cells began to attain a more recognizable, columnar and polarised epithelial morphology. Cells grown on a type-I collagen gel which was free-floating showed a selective expression of alkaline phosphatase at the apical surfaces of approximately 10% of the cells. This expression was detectable by electron microscope histochemistry but could not be detected biochemically. Treatment of MAC 15j cells grown on a released collagen matrix with tetramethyl-urea (20mM) accelerated the expression of alkaline phosphatase activity at the apical surface as detected by microscopy.

A new method for the 3‐D in vitro growth of human RT112bladder carcinoma cells using the alginate culture technique

“…In the case of 2-D cell culture, contact between RTll2 cells and lens capsules markedly influenced the growth behaviour of these cells, while artificial supports hindered the epithelial morphogenesis and the development of the epithelial cell polarity. Confirming the findings of others [52,71] we can support the suggestion that the degree of morphological and physiological differentiation of epithelial cells is largely affected by the physical constitution of the substratum as well as by factors of their local environment [18,63]. In addition, more recent ihvestigations have demonstrated that ceils cultured on a natural kidney capsule revealed a morphology of the tight junctions almost identical to that of native cells while ceils grown on plastic or on collagen coatings expressed only an irregular pattern of junctional strands [33].…”

“…When grown on solid substrata the cells are forced to feed completely from the apical side whereas permeable filters permit access of nutrients to both surfaces, thus promoting polarized growth of the cells [13,28,71]. The basal lamina and some of its individual components are important factors for the expression of the polarized phenotype of epithelial cells in vitro [40,54,67].…”

“…The basal lamina and some of its individual components are important factors for the expression of the polarized phenotype of epithelial cells in vitro [40,54,67]. Several previous works have reported a strong influence of the extracellular matrix on various epithelial activities [ 1,6,18,22,30,31,57,64,71]. The lens capsule represents the natural lamina densa of the basal lamina [7] and we, therefore, expected that the culture of RT112 cells on this material would lead to both a prismatic phenotype and a higher cell density compared with the cell culture on solid, artificial supports.…”

Interferon‐γ Tempers the Expression of Carcinoembryonic Antigen Family Molecules in Human Colon Cells: a Possible Role in Innate Mucosal Defence

“…These had internal microvillous surfaces and content that was positive for all examined CEA subfamily antigens except CEACAM7. These cytoplasmic neostructures have been found in a variety epithelial neoplasm [39] and cancer cell lines, including HT‐29 cells [57, 58]. It is believed that, in epithelial cells, the appearance of the intracellular lumina is connected with a dysfunction of the Golgi complex caused by the loss of cell polarity in cases of malignancy or cell isolation.…”