The Isolation of Living Cells from Animal Tissues

Rinaldini, Luis M.

doi:10.1016/s0074-7696(08)62696-0

Cited by 143 publications

(25 citation statements)

References 190 publications

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“…A similar phenomenon seems to have been observed in the classical research in experimental embryology in the 1950s (Auerbach & Grobstein, 1958;Rinaldini, 1958;Moscona, 1961;Steinberg, 1963). Auerbach and Grobstein (1958) reported the appearance of 'gummy material' which trapped single cells and small cell masses when they attempted to prepare free cells from kidney rudiment by treatment with trypsin or chymotrypsin.…”

supporting

confidence: 73%

Serine protease-induced enhancement of blood-borne metastasis of rat ascites tumour cells and its prevention with deoxyribonuclease

Sugihara

Yamamoto²,

Tsuruta³

et al. 1990

Br J Cancer

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Summary Serine proteases, such as a-chymotrypsin or elastase, caused an aggregation of rat ascites tumour cell lines, AH-130, AH-109A and YS, in a protein free medium which preserved the cell viability. This aggregation, which was monitored spectrophotometrically, was dependent upon the protease activities and was resistant to treatment with either a calcium chelating reagent (EDTA) al., 1973;Liotta et al., 1980;Sloane et al., 1982;Wang et al., 1980). It has also been shown that an important event in blood-borne metastasis is the intercellular adhesion and aggregation of circulating tumour cells (Nicolson & Winkelhake, 1975). The aggregated state of tumour cells in the microcirculation may be favourable to lodgement in distant organs. In this sense, the role of the plasma clotting system (Kinjo, 1978) or platelets (Hara et al., 1980) in blood-borne metastasis has been emphasised with special reference to the formation of floating or plugged masses of tumour cells.In our preliminary experiments, serine proteases such as a-chymotrypsin or elastase caused an aggregation of rat ascites tumour cell lines, AH-130, AH-109A and YS, without any appreciable change in cell viability. In the present study, we have examined a potential augmenting effect of proteaseinduced cell aggregation in vitro and in vivo on blood-borne metastasis using the lung-colonisation model with rat ascites tumour cells. In addition, we also examined the effectiveness of DNase I treatment (which prevented the protease-induced cell aggregation) on blood-borne metastasis in the lung. Materials and methods AnimalsFemale Donryu rats weighing 130-180 g were generally used in this study. These rats received a standard rat pellet diet and tap water ad lib. ReagentsCrystallised bovine pancreatic a-chymotrypsin, elastase, neuraminidase, bovine muscular actin, bovine pancreatic DNase I and calf thymus DNA were purchased from Sigma Chemical Co. (St Louis, MO, USA). Chymostatin and elastatinal were purchased from Peptide Institute Inc. (Osaka, Japan). Hanks' balanced salt solution (HBSS) (without phenol red) was purchased from Nissui Pharm. Co. (Tokyo, Japan), HBSS (without Ca2 , Mg2 + and phenol red) from Gibco (Grand Island, NY, USA). An assay kit for lactate dehydrogenase (LDH) was obtained from Shinotest Laboratories (Kanagawa, Japan).The DNase I was further purified using a high performance liquid chromatography system (LKB) with a TSK gel 3,000 SW gel permeation column (Tosoh). The final preparation was pure as judged by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate with or without 1% P-mercaptoethanol.

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supporting

confidence: 73%

Serine protease-induced enhancement of blood-borne metastasis of rat ascites tumour cells and its prevention with deoxyribonuclease

Sugihara

Yamamoto²,

Tsuruta³

et al. 1990

Br J Cancer

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“…The first factor, the effective Na+-conductivity of the surface gNa, appears to be quite sensitive to the specific nature, chemical structure and intercellular associations of the cell surface polymers, properties which in turn are determined by the particular metabolic expression of the cell and the immediate molecular environment. O f pardcular interest here are the mucopolysaccharides and similar saccharide polymers (collectively denoted herein as MPS) known to be associated with the cell surface [Rinaldini, 1958;Brandt, 1958;Bell, 1962;D orfman, 1963]. For example, the sur face MPS appear to be a primary functional agent in immunological expression [Moscona and Moscona, 1952;D avies, 1959], and it is well established that immunologically active changes in the cell sur face accompany malignant transformation in cells [Alexander, 1966;Ambrose, 1966]; this surface change accompanying malignant trans formation has also been demonstrated by electrophoretic mobility studies [Ambrose, 1966].…”

Section: E M-metabolic Feedback Circuits and Mitosis Controlmentioning

confidence: 99%

“…Trypsinization of living tissue has long been used as a method for preparation of free cell suspensions [Rous and J ones, 1916;Rinaldini, 1958]. Trypsin digestion apparently breaks protein-associated bonds in the cell surface molecular complex which are directly or indirectly responsible for binding the cells together.…”

Section: Itotic Stimulation By Cell Surface Treatmentsmentioning

confidence: 99%

Variation of the Transmembrane Potential Level as a Basic Mechanism of Mitosis Control

Cd¹

1970

Oncology

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A range of experimental observations suggests that a significant correlation exists between the level of the electrical transmembrane potential difference in somatic cells and the degree of their mitotic activity. The present paper, after review of pertinent experimental background data, assumes that a functional relationship between potential level and mitotic activity does exist and, invoking the precepts of classical membrane potential theory, proceeds with the formulation of a basic theory of mitosis control wherein the intracellular ionic conditions associated with various levels of the potential difference act to regulate DNA synthesis and other essential preparations for mitosis. The theory links the activity of the potential-generation mechanisms of the cell surface complex, and hence mitogenic activity, with cellular metabolism and with external environmental influences through an explicit system of interacting feedback circuits. Inherent in the overall theoretical development is the formulation of a unified theory of the cytogenetic etiology and maintenance of the malignant state. Additional specific experimental evidence is cited in support of the theoretical concepts developed.

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“…The preparation of cell suspensions by treatment of tissue with proteolytic enzymes is a well established technique in the field of tissue culture (Rinaldini, 1958). The origin of cell adhesiveness is itself not understood, some workers supporting the suggestion that an intercellular cement exists (Moscona, 1962(Moscona, , 1968Kuroda, 1968), the cement possibly being of proteinaceous material, while others believe that cell interaction is governed by a non-specific balancing of attractive forces, governed by 252 EPITHELIAL GROWTH FACTOR the properties of cell surfaces and the viscosity of intercellular material (Curtis, 1970;Curtis & de Vyver, 1971).…”

Section: Discussionmentioning

confidence: 99%

Protein fractions from mouse salivary glands affecting epithelial cells in organ culture

Banks¹,

Walter²

1973

The Journal of Physiology

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The Isolation of Living Cells from Animal Tissues

Cited by 143 publications

References 190 publications

Serine protease-induced enhancement of blood-borne metastasis of rat ascites tumour cells and its prevention with deoxyribonuclease

Serine protease-induced enhancement of blood-borne metastasis of rat ascites tumour cells and its prevention with deoxyribonuclease

Variation of the Transmembrane Potential Level as a Basic Mechanism of Mitosis Control

Protein fractions from mouse salivary glands affecting epithelial cells in organ culture

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