ESTABLISHMENT AND CRYOPRESERVATION OF FIBROBLAST CELL LINE FROM A SUMATRAN RHINOCEROS (Dicerorhinus sumatrensis)

AbstractThe number of biomaterials used in biomedical applications has rapidly increased in the past two decades. Fluorapatite (FA) is one of the inorganic constituents of bone or teeth used for hard tissue repairs and replacements. Fluor-hydroxyapatite (FHA) is a new synthetically prepared composite that in its structure contains the same molecular concentration of OH− groups and F− ions. The aim of this experimental investigation was to use the embryonal mouse fibroblast cell line NIH-3T3 for comparative study of basal cytotoxicity of fluoridated biomaterials FHA and FA discs. Hydroxyapatite (HA) disc, high-density polyethylene as negative control and polyvinyl chloride (PVC) containing organotin stabilizer as positive control were used as standard biomaterials. The appropriateness of the use of NIH-3T3 cells and their sensitivity for tested biomaterials were evaluated on the basis of five cytotoxic end points: cell proliferation, cell morphology, lactate dehydrogenase (LDH) released, protein and DNA cell content. The basal cytotoxicity of FHA, FA and HA discs was measured by direct contact method. FHA composite, FA and HA demonstrated in cell line NIH-3T3 nearly similar basal cytotoxicity increasing with the time of treatment. After 72 h of biomaterials treatment, about 25% inhibition of cell number, unchanged morphology of dividing cells, 6.31–0.16% increase of released LDH, about 10% inhibition of cell protein content and about 20% inhibition of DNA content was found. On the other hand, from the growth rates it resulted that NIH-3T3 cells, affected by tested biomaterials, divided about 20% slowlier than the control (untreated cells). Using the linear regression analysis we found out that deviations in measurements of cytotoxicity by four methods were as follows: less than 10% for cell number, protein and DNA content methods and 12.4% for released LDH method. Based on a good correlation of the cytotoxicity of biomaterials obtained from all end points we could conclude that fibroblast NIH-3T3 cell line was appropriate for measuring the basal cytoxicity of tested biomaterials.

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Type I and type II interferons up-regulate functional type I interleukin 1 receptor in a human fibroblast cell line TIG-1

Cytokine ◽

10.1016/1043-4666(94)90182-1 ◽

1994 ◽

Vol 6 (5) ◽

pp. 555

Author(s):

Takemasa Takii ◽

Hidetoshi Hayashi ◽

Kikuo Onozaki

Keyword(s):

Cell Line ◽

Human Fibroblast ◽

Interleukin 1 ◽

Fibroblast Cell ◽

Type I ◽

Type Ii ◽

Fibroblast Cell Line ◽

Functional Type ◽

Human Fibroblast Cell Line ◽

Human Fibroblast Cell

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Rosette formation by a mouse fibroblast cell line

Nature ◽

10.1038/248514a0 ◽

1974 ◽

Vol 248 (5448) ◽

pp. 514-515 ◽

Cited By ~ 7

Author(s):

E. V. ELLIOTT ◽

R. S. KERBEL ◽

B. J. PHILLIPS

Keyword(s):

Cell Line ◽

Fibroblast Cell ◽

Mouse Fibroblast ◽

Rosette Formation ◽

Fibroblast Cell Line ◽

Mouse Fibroblast Cell ◽

Mouse Fibroblast Cell Line

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Activation and repression of a beta-globin gene in cell hybrids is accompanied by a shift in its temporal replication

Molecular and Cellular Biology ◽

10.1128/mcb.9.8.3524-3532.1989 ◽

1989 ◽

Vol 9 (8) ◽

pp. 3524-3532

Author(s):

V Dhar ◽

A I Skoultchi ◽

C L Schildkraut

Keyword(s):

Cell Line ◽

Hybrid Cell ◽

Globin Gene ◽

Fibroblast Cell ◽

Globin Genes ◽

Fibroblast Cell Line ◽

Beta Globin ◽

Hybrid Cells ◽

Beta Globin Gene ◽

Cell Hybrids

To investigate whether a switch in the transcriptional activity of a gene is associated with a change in the timing of replication during the S phase, we examined the replication timing of the beta-globin genes in two different types of somatic cell hybrids. In mouse hepatoma (Hepa 1a) x mouse erythroleukemia (MEL) hybrid cells, the beta-globin gene from the MEL parent is transcriptionally inactivated and is later replicating than in the parental MEL cell line. In human fibroblast (GM3552) x MEL hybrid cells, the human beta-globin gene is transcriptionally activated, and all of the sequences within the human beta-globin domain (200 kilobases) we have examined appear to be earlier replicating than those in the parental fibroblast cell line. The chromatin configuration of the activated human beta-globin domain in the hybrids is relatively more sensitive to nucleases than that in the fibroblasts. Furthermore, major nuclease-hypersensitive sites that were absent in the chromatin flanking the distal 5' region of the human beta-globin gene cluster in the parental fibroblast cell line are present in the transcriptionally activated domain in the hybrid cell line. These results suggest that timing of replication of globin genes has been altered in these hybrid cells and thus is not fixed during the process of differentiation.

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Oncogene-induced transformation of a rat embryo fibroblast cell line is enhanced by tumor promoters

Molecular and Cellular Biology ◽

10.1128/mcb.6.6.1943-1950.1986 ◽

1986 ◽

Vol 6 (6) ◽

pp. 1943-1950

Author(s):

W L Hsiao ◽

T Wu ◽

I B Weinstein

Keyword(s):

Cell Line ◽

Fold Increase ◽

Fibroblast Cell ◽

Tumor Promoters ◽

Fibroblast Cell Line ◽

Ras Oncogene ◽

Rat Embryo ◽

Synergistic Interactions ◽

Multistage Carcinogenesis ◽

System Tumor

Rat embryo fibroblast cell line 6 was transfected with plasmid pT24, which contains the activated human bladder c-Ha-ras oncogene, and the cells were grown continuously in the absence or presence of the tumor promoters 12-O-tetradecanoyl phorbol-13-acetate (TPA) or teleocidin. The presence of TPA or teleocidin led to a 6- to 14-fold increase in the number of morphologically transformed foci. No transformed foci were seen when rat 6 cells were transfected with the normal c-Ha-ras oncogene in the absence or presence of TPA, or in cells simply treated with TPA or teleocidin. Enhancement of pT24-induced foci was seen even when the addition of TPA was delayed until day 16. In transfection studies with the drug resistance genes gpt and neo, TPA and teleocidin did not increase the number of Gpt+ or Neo+ colonies. When rat 6 cells were cotransfected with pT24 and neo genes and grown in the absence or presence of TPA, the presence of TPA did not increase the yield of Neo+ colonies but caused a fivefold increase in the number of Neo+ colonies that displayed a transformed morphology. Southern blot analyses of DNAs obtained from these clones indicated that TPA treatment did not influence the extent of integration of either the pT24 or neo gene. DNA samples from all of the morphologically transformed cells displayed a characteristic 2-kilobase SacI fragment homologous to pT24 DNA and expressed relatively high levels of the corresponding mRNA. Our findings indicate that in this system tumor promoters do not simply enhanced the process of DNA transfection per se. Thus, this model system may be useful for analyzing synergistic interactions between tumor promoters and activated oncogenes during multistage carcinogenesis. It may also serve as a simple screening test for detecting new tumor promoters.

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