A New In-Vitro Model to Study Tumour Cell Invasion

Tchao, Ruy; Schleich, Annelies; Frick, Manfred; Mayer, August

doi:10.1007/978-94-009-8925-2_7

Cited by 4 publications

(5 citation statements)

References 5 publications

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“…Studies based on the human amniotic membrane have focused primarily on the invasive and degradative processes involved in tumor cell interaction with an ECM, since it is considered to be one of the more discernible (measurable) aspects of the invasion process. These studies have provided helpful information pertaining to the morphological and mechanical aspects of cell invasion [16,18,34,37,40], as well as to the underlying mechanisms involved in this intricate cascade of events. It is highly probable that the murine melanoma tumor cells used in this study produce d specific collagenases [19,36,44] in addition to plasminogen activator [21,28,36], a serine protease, to traverse the BM, rather than to rely on simple haptotaxic migration [15,22].…”

Section: Discussionmentioning

confidence: 99%

“…To investigate the invasive characteristics and associated properties of tumor cells, numerous innovative models have been presented [for review, see 18]. One of the more popular in vitro techniques described is the amniotic BM model [15,40]. This particular model offers a number of advantages, including: (1) a large quantity of material which can be isolated from each human placental sample; (2) a membrane and connective tissue stroma with an ideal composition for studying the degradative interactions of tumor cells with an ECM, which is identical to an in vivo environment; and (3) the ease of conducting morphological, immunohistochemical, and biochemical studies in this system.…”

Section: Introductionmentioning

confidence: 99%

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In vitro quantification of melanoma tumor cell invasion

et al. 1985

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In order to quantify the invasiveness of melanoma tumor cells in vitro, a modification of the amniotic basement membrane (BM) model, described by Liotta et al. (Cancer Letters, 11, 141, 1980), was used in combination with radiolabeled tumor cells. B16-F10 metastatic murine melanoma cells and a derived clone (B16-F10L) were prelabeled with 0.1 muCi/ml of [14C]thymidine for 20-24 h in serum-free medium at 37 degrees C. Following incubation, fetal bovine serum was added to a concentration of 5 per cent, and the cells were allowed to grow to confluency for the next 24-28 h. The labeled cells were seeded onto amniotic membranes situated in Membrane Invasion Culture System (MICS) chambers at a density of 2.5 X 10(4) per well. At various times points, radioactivity of tumor cells that completely traversed the membrane was determined using an under-the-membrane sampling method. The average percent invasion demonstrated by the B16-F10 line was 2.75 per cent, and 3.65 per cent exhibited by the B16-F10L cell line after 48-53 h in vitro. Since it was apparent that some variability in thickness existed among membrane samples, a morphological analysis was performed on five sectors of a three-inch-diameter sample from four different placentae. Differences and similarities in BM thickness within the same sector were noted by this technique and could possibly contribute to some variability observed in tumor cell invasion in this model. Another parameter examined was the proliferation of tumor cells in the upper and lower wells of the MICS chambers. By 48 h, approximately 32.1 per cent of the B16-F10 cell line as well as the clone had replicated in the upper wells associated with the BMs compared with a 32.9 per cent replication in the lower wells, which reaffirmed the viability of the tumor cells under experimental conditions and insured similarly replicating populations of cells. In order to quantify the invasiveness of radiolabeled tumor cells accurately through a biological membranous barrier, the proper concentration of cells must be used, tumor cell heterogeneity should be taken into consideration, the technique of sampling radiolabeled invasive cells should be critically analysed, and thickness of the membranous barrier should all be considered as possible important factors in the quantitative analyses.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In vitro quantification of melanoma tumor cell invasion

et al. 1985

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“…The in vitro assay for tumor cell invasion of human placental amnion has been described [2,16,30,33,40]. Briefly: amnion was dissected from fresh placenta and treated with 0-1 N ammonium hydroxide for 30min to remove epithelia.…”

Section: Invasion Assaymentioning

confidence: 99%

“…The present study describes a novel method for the selection of invasive tumor cells using a variation of the in vitro human amnion assay [2,16,30,33,40,43]. The assay was modified to require cellular invasion of basement membrane and stroma in vivo, where tumor-host interactions can influence this process.…”

Section: Introductionmentioning

confidence: 99%

A novel method for selection of invasive tumor cells: Derivation and characterization of highly metastatic K1735 melanoma cell lines based onin vitro andin vivo invasive capacity

et al. 1988

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Tumor cell invasion of basement membranes is required at several steps in the process of metastasis. To study the genetic and biochemical events mediating invasion, a variant cell line (TK) was selected from the metastatic M2 K1735 murine melanoma cell line. A novel selection procedure was used, based on in vitro and in vivo invasion and growth upon basement membrane and stroma. Additionally, two extrapulmonary metastases of the TK cell line, TK-Eve and TK-Liver, were established as cell lines and characterized. The TK cell line demonstrates greater metastatic potential in vivo and invasive ability in vitro than the parent M2 cell line, confirming the validity of the selection procedure. In addition, the M2 and TK cell lines were examined for other cell functions involved in the metastatic process. Cellular growth rates and sensitivity to T lymphocyte and natural killer cell lysis were not determining factors in the metastatic potentials of the M2 and selected cell lines; possible macrophage contribution to metastatic behavior was noted. [35S]methionine pulse labeling of protein synthesis and karyotypic analysis confirm the close relationship of parental and selected cell lines.

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“…Several days later, after outgrowth from the normal inocula was luxuriant, tumor cells were added, usually as a cell line growing in a fragment of Gelfoam. Not recognized at the time was that our model had the same deficiency in regard to gradients as other organ culture models where tumor is placed in surface contact with a rounded organ culture of normal tissue such as chick embryo heart (33), or on a biologic membrane such as chick chorioallantoic membrane (34,35), or human amnion (36,37). In all of these, when invasion occurs the tumor cells must move away from the source of renewal of metabolites, i.e.…”

Section: Gradients and Neoplastic Blockadementioning

confidence: 99%

The propagation of cancer, a process of tissue remodeling

Leighton¹,

Tchao²

1984

Cancer Metast Rev

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Effective study of the malignant phenotype at the tissue level requires model systems that are intelligible both to cell biologists and to pathologists, and that also observe the spatial imperatives intrinsic to tissues in nature. Malignant cells commonly appear in multicellular units, and growth of tumor tissue is seen as an increase in the number of cells and multicellular units. The supporting stroma frequently has an abnormal appearance, and this component of the tissue also increases in mass as the tumor enlarges and spreads. The direction of invasion is influenced by the direction of available metabolites. Histophysiologic gradient culture complies with nature's spatial rationale, since at the substrate-parenchymal interface three functions coincide. These are anchorage, initiation of epithelial renewal, and complete exchange of metabolites. Our model system provides a setting for reconstructing and manipulating many features of the malignant phenotype seen in cancer tissues in nature, such as abnormalities in the sequence of maturation of stratified epithelium, hyperplasias, dysplasias, interaction between different types of epithelium, aggregate formation, tumor angiogenesis, and neoplastic blockade.

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A New In-Vitro Model to Study Tumour Cell Invasion

Cited by 4 publications

References 5 publications

In vitro quantification of melanoma tumor cell invasion

In vitro quantification of melanoma tumor cell invasion

A novel method for selection of invasive tumor cells: Derivation and characterization of highly metastatic K1735 melanoma cell lines based onin vitro andin vivo invasive capacity

The propagation of cancer, a process of tissue remodeling

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