Cytochemical analysis of vero cells on type I collagen gels in long-term culture

Maria, Silvya Stuchi; Wada, Maria Lúcia Furlan

doi:10.1007/s11626-997-0152-9

Cited by 11 publications

(8 citation statements)

References 17 publications

(13 reference statements)

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“…This behavior is similar to the normal growth pattern on nonbiological surfaces, such as a glass coverslip (20–22). On the other hand, it is different from growth patterns on a biological substratum, such as a three‐dimensional Type I collagen gel (23) or dry collagen I sponges (24). The cells respond differently to substratum characteristics such as its composition, consistency, and flexibility (25).…”

Section: Discussionmentioning

confidence: 99%

Vero Cell Growth and Differentiation on Poly(l‐Lactic Acid) Membranes of Different Pore Diameters

Santos

Barbanti²,

Duek³

et al. 2001

Artificial Organs

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In the last few years, the demand has increased for research on polymeric materials, which can be used as substitutes for injured tissues and organs or to improve their regeneration. In this work, we studied poly(L-lactic acid) (PLLA) membranes, a resorbable biomaterial, which were either dense or had different pore diameters (less than 45 microm, between 180 and 250 microm, and between 250 and 350 microm), in relation to stimulation of cell adhesion, growth, and differentiation in vitro. We used Vero cells, a fibroblastic cell line, as the biological model of investigation. We found that cells attached slowly to all PLLA membranes studied. On the other hand, once the adhesion occurs, the cells are able to grow and differentiate on the different polymers. The cells grew to form a confluent monolayer and were capable of producing collagen Type IV and fibronectin on different PLLA membranes. This behavior indicates that cells try to create a better environment to stimulate their growth. This also indicates that Vero cells alter their differentiation pattern once they are producing extracellular matrix molecules related to epithelial differentiation.

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Section: Discussionmentioning

confidence: 99%

Vero Cell Growth and Differentiation on Poly(l‐Lactic Acid) Membranes of Different Pore Diameters

Santos

Barbanti²,

Duek³

et al. 2001

Artificial Organs

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“…These cells were grown in Dulbecco's modified Eagle's minimum essential (DMEM) medium supplemented with 10% fetal bovine serum (FBS), 50 U/ml penicillin and 50 µg/ml streptomycin and maintained at 37°C under 5% CO 2 atmosphere. Type 1 collagen gel, extracted from rat‐tail tendons, was prepared as previously described [Maria and Wada, 1997] with minor modifications. For growth curves, 1 × 10 4 cells of both A172 and T98G cell lines were plated in 24‐wells plates onto either plastic or collagen (175 µl/well, 2.5 mg/ml).…”

Section: Methodsmentioning

confidence: 99%

Downregulation of the RECK‐tumor and metastasis suppressor gene in glioma invasiveness

Corrêa

Brohem

Winnischofer

et al. 2006

J of Cellular Biochemistry

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Invasive behavior is the pathological hallmark of malignant gliomas, being responsible for the failure of surgery, radiation, and chemotherapy. Matrix metalloproteinases (MMPs) are essential for proper ECM remodeling and invasion. The tumor and metastasis suppressor RECK protein regulates at least three members of the MMPs family: MMP-2, MMP-9, and MT1-MMP. In order to mimic the in vivo invasion process, A172 and T98G, respectively, non-invasive and invasive human glioblastoma cell lines, were cultured onto uncoated (control) or type I collagen gel-coated surface, and maintained for up to 7 days to allow establishment of the invasive process. We show that the collagen substrate causes decreased growth rates and morphological alterations correlated with the invasive phenotype. Electronic transmission microscopy of T98G cells revealed membrane invaginations resembling podosomes, which are typically found in cells in the process of crossing tissue boundaries, since they constitute sites of ECM degradation. Real time PCR revealed higher RECK mRNA expression in A172 cells, when compared to T98G cells and, also, in samples obtained from cultures where the invasive process was fully established. Interestingly, the collagen substrate increases RECK expression in A172 cells and the same tendency is displayed by T98G cells. MMPs-2 and -9 displayed higher levels of expression and activity in T98G cells, and their activities are also upregulated by collagen. Therefore, we suggest that: (1) RECK downregulation is critical for the invasiveness process displayed by T98G cells; (2) type 1 collagen could be employed to modulate RECK expression in glioblastoma cell lines. Since a positive correlation between RECK expression and patients survival has been noted in several types of tumors, our results may contribute to elucidate the complex mechanisms of malignant gliomas invasiveness.

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“…Scanning Electron Microscopy (SEM) -Particles of MBB were adhered to glass coverslips coated with three-dimensional rat tail hydrated type I collagen (13) to avoid displacement of the MBB granules. Balb/c 3T3 fibroblasts (clone A31) were plated onto the MBB at the density of 5x10 4 cells/coverslips.…”

Section: Morphologic Analysismentioning

confidence: 99%

Evaluation of the cytocompatibility of mixed bovine bone

et al. 2007

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Treatment of bovine bone with peroxides and chaotropic agents aims to obtain an acellular bone matrix that is able to maintain the collagen-apatite complex and a higher mechanical resistance, a mixed biomaterial hereby named mixed bovine bone (MBB). The purpose of this study was to evaluate the cytocompatibility of MBB and cell-MBB interaction. Cell morphology, number of viable cells, ability to reduce methyltetrazolium and to incorporate neutral red upon exposure to different concentrations of the hydrosoluble extract of MBB were assessed in Balb-c 3T3 cells according to ISO 10993-5 standard. The interaction between cells and MBB surface was evaluated by scanning electron microscopy. The water-soluble MBB extracts were cytotoxic and led to cell death possibly due to its effect on mitochondrial function and membrane permeability. Cells plated directly onto the MBB did not survive, although after dialysis and material conditioning in DMEM + 10% FCS, the cells adhered and proliferated onto the material. It may be concluded that, in vitro, water-soluble MBB extracts were cytotoxic. Nevertheless, MBB cytotoxic effect was reverted by dialysis resulting in a material that is suitable for cell based-therapy in the bioengineering field.

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Cytochemical analysis of vero cells on type I collagen gels in long-term culture

Cited by 11 publications

References 17 publications

Vero Cell Growth and Differentiation on Poly(l‐Lactic Acid) Membranes of Different Pore Diameters

Vero Cell Growth and Differentiation on Poly(l‐Lactic Acid) Membranes of Different Pore Diameters

Downregulation of the RECK‐tumor and metastasis suppressor gene in glioma invasiveness

Evaluation of the cytocompatibility of mixed bovine bone

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