Estudo comportamental da linhagem celular Vero quanto cultivada em substrato de gel de colageno tipo I

AUTOR(ES)

Silvya Stuchi Maria

DATA DE PUBLICAÇÃO

1994

RESUMO

Collagen is widely employed as substratum for cell adhesion and proliferation. Collagen-based substrata may be prepared in different ways, as dry films, floating gel or three dimensional structures. Many cell types undergo morphophysiological alterations when placed on different collagen substrata. Not only the presence but also the organization of individual components of the extracellular matrix are important for cells to assume a given phenotype. Another aspect about the behavior of cultured cells is their ability to contract collagen gels, similarly to process occurrings in the dermis during wound healing, in fibrosis, and in connective tissue development .We have here studied the behavior of Vero cells grown on type I collagen gels. Samples were harvested afier 1, 5, 10, 15, 20, 25 e 30 days of incubation, fixed in Kamovsky s fixative and embedded in Paraplast Plus. Sections 5 f-tm thick were obtained, dewaxed and stained with either hematoxilin-eosin (HE), toluidine blue at pH 4,0 (AT), xylidine ponceau (XP) or PAS. Parallel experiments were conducted to evaluate the effect of foetal bovine se rum on the cell s ability to contract collagen gels. The cells first adhered to the gel surface and constitute a monolayer afier 24 hours of culturing. The cells were initially rounded but became fibroblast-like and showed intense nuclear and cytoplasmatic metachromasy. Reactivity with XP was observed in the nuclei and cytoplasm and P AS-positive material accumulated in the cytoplasm. Mer 5 days, the cells were fibroblast-like and grew in multiple layers. Foci of cells invading the gel were observed at this time and numerous metachromatic granules were deposited in the gel. These granules were also XP and P AS positive. Longer periods of cell growth resulted in enhanced proliferation and invasion of the gel, but the staining characteristics were preserved. At 20 days, the cells occupied a large extension of the gel, which showed aspects of loose connective tissue. On the 25th. day, the gel surface is highly folded and XP­ stained collagen fibers were mostly aligned featuring the contraction of the gel by Vero cells.showing contraction of the gel by Vero cells. The cells inside the gel presented aspects of senescense, with cytoplasm fragmentation and nuclear picnosis. These aspects were aIs o observed on the 30th.day. In the presence of serum, the cells contracted the gel in 10% and 50% of its previous length after 1 and 5 days, respectively. Without serum the values were 7% and 15% shorter, after 1 and 5 days, respectively.The results demonstrated that Vero cells adhere to, proliferate and invade collagen gels in vitro characterizes a substratum-induced differentiation processo The cytochemical characteristics of the granules entrapped in the gel indicate that they are constituted by sulphated and/or carboxylated extracellular glycoproteins produced and secreted by the cells. It was also shown that the contraction of the gel is a serum-dependent activity of Vero cells

ASSUNTO(S)

linhagem (genetica) colageno celulas - transformação

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