Rapid melanization of bomirski amelanotic melanoma cells in cell culture

Transfer of Bomirski amelanotic melanoma ceils from in vivo to in vitro growth conditions results in occurrence of rapid melanization in their cytoplasm. The melanized ceils from primary cell culture initiate tumours in hamsters, which do not contain traces of melanin and resemble typical amelanotic melanoma.

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Do alterations in gene expressions influence tumorigenesis in the transmissible venereal tumor in dogs?

Ciência Rural ◽

10.1590/0103-8478cr20200082 ◽

2020 ◽

Vol 50 (11) ◽

Author(s):

Haline Ballestero Fêo ◽

Luis Mauricio Montoya Flórez ◽

Ricardo Seiti Yamatogi ◽

Anderson do Prado Duzanski ◽

João Pessoa Araújo Junior ◽

...

Keyword(s):

Cell Culture ◽

Tumor Suppressor Genes ◽

Primary Cell Culture ◽

Gene Mutations ◽

Tp53 Gene ◽

Primary Cell ◽

Loss Of Function ◽

Gene Expressions

ABSTRACT: Canine transmissible venereal tumor (CTVT) is a transmissible neoplasm, which spreads naturally between dogs through the halogenic transfer of tumor cells, mainly during coitus. It is the oldest known tumoral lineage in nature and reports on gene mutations have been extended. Also, this tumor shares several genetic mutations with some cancers in humans, among them lung carcinomas, melanoma, prostate, breast, among other cancers. Thus, expression of tumor suppressor genes such as TP53, P21, and apoptosis-related genes such as BAX, BCL-2, and BCL-xL, both in vivo and in vitro (primary cell culture) were quantified. In the present study, the comparison of gene expression, the TP53 gene, in most cases, was shown to be high in the majority of tissues (65%) and primary cell culture (100%), while BCL-2, BCL-xL, and BAX presented variation among the animals analyzed. Moreover, in these situations, the results suggested that the apoptotic regulation of these genes did not occur for TP53. The P21 gene was shown to be mostly normal (70%); although, absence (6%) and underexpressions (24%) were also observed. Statistical analysis of the BCL-xL gene demonstrated significant differences between the tissues of the animals when compared to the cell cultures; however, to the other genes, no statistical difference was observed between the groups. Preliminarily, the results suggested the presence of alterations in the gene expressions of the TP53, P21, BAX, BCL-2 and BCL-xL leading to loss of function in these genes, which affect the tumorigenesis of CTVT.

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Some properties of Bomirski Ab amelanotic melanoma cells, which underwent spontaneous melanization in primary cell culture

Journal of Cancer Research and Clinical Oncology ◽

10.1007/bf01884251 ◽

1985 ◽

Vol 109 (1) ◽

pp. 29-37 ◽

Cited By ~ 19

Author(s):

Andrzej Słomiński

Keyword(s):

Cell Culture ◽

Primary Cell Culture ◽

Melanoma Cells ◽

Amelanotic Melanoma ◽

Primary Cell

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Expression of Selected Connexin and Aquaporin Genes and Real-Time Proliferation of Porcine Endometrial Luminal Epithelial Cells in Primary Culture Model

BioMed Research International ◽

10.1155/2020/7120375 ◽

2020 ◽

Vol 2020 ◽

pp. 1-15 ◽

Cited By ~ 1

Author(s):

Katarzyna Wojtanowicz-Markiewicz ◽

Magdalena Kulus ◽

Sandra Knap ◽

Ievgenia Kocherova ◽

Maurycy Jankowski ◽

...

Keyword(s):

Cell Culture ◽

Epithelial Cells ◽

Primary Cell Culture ◽

Water Movement ◽

Inorganic Ions ◽

Primary Cell ◽

Lag Phase ◽

Luminal Epithelial Cells

Luminal epithelial cells are the first embryonic-maternal contact site undergoing very specific changes associated with reproductive processes. Cells prepare for embryo development by increasing their volume, with the help of aquaporins that provide a transcellular path of rapid water movement during the secretion and absorption of fluids, as well as connexins enabling the flow of inorganic ions and small molecules. In this work, we have examined how AQPs and Cx’s behave in luminal epithelium primary cell culture. Cells obtained from porcine specimen during slaughter were primarily in vitro cultured for 7 days. Their proliferation patterns were then analyzed using RTCA, with the expression of genes of interest evaluated with the use of immunofluorescence and RT-qPCR. The results of these changes of gene of interest expression were analyzed on each of the seven days of the porcine luminal primary cell culture. Our study showed that the significant changes were noted in the case of Cx43, whose level of protein expression and distribution increases after 120 hours of culture, when the cells enter the lag phase, and maintains an upward trend until the end of the culture. We noted an increase in AQP4, AQP7, AQP8, and AQP11 levels throughout the entire culture period, while the largest differences in expression were found in AQP3, AQP4, and AQP10. The obtained results could become a point of reference for further in vivo and clinical research. Experiments conducted with these proteins showed that they influence the endometrial fluid content during the oestrous cycle and participate in the process of angiogenesis, which intensifies during endometrial development.

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Tyrosinase activity in primary cell culture of amelanotic melanoma cells

Bioscience Reports ◽

10.1007/bf01121029 ◽

1983 ◽

Vol 3 (11) ◽

pp. 1027-1034 ◽

Cited By ~ 10

Author(s):

A. Słomiński ◽

P. W. D. Scisłowski ◽

A. Bomirski

Keyword(s):

Gel Electrophoresis ◽

Primary Cell Culture ◽

Soluble Fraction ◽

Calf Serum ◽

Melanoma Cells ◽

Amelanotic Melanoma ◽

Tyrosinase Activity ◽

Logarithmic Phase

After transfer of the Ab amelanotic melanoma cells from in vivo to in vitro growth conditons tyrosinase activity in their soluble fraction rapidly increased. This increase lasted to the middle of the logarithmic phase of growth and was followed by a decrease of tyrosinase activity, which was accompanied by accumulation of melanin in the cells. Calf serum stimulated simul-taneously tyrosinase activity, melanin synthesis, and proliferation of the melanoma ceils. Acrylamide-gel electrophoresis patterns of soluble tyrosinase from the Ab melanoma cells cultured in vitro consisted of two bands, similarly as soluble tyrosinase from the Ma melanotic melanoma cells freshly isolated from solid tumors.

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In Vivo-like model of glial tumors: Creation of primary cell lines.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.15_suppl.e14515 ◽

2020 ◽

Vol 38 (15_suppl) ◽

pp. e14515-e14515

Author(s):

Sofia V. Timofeeva ◽

Oleg I. Kit ◽

Anastasia O. Sitkovskaya ◽

Irina V. Mezhevova ◽

Svetlana Yu. Filippova ◽

...

Keyword(s):

Cell Culture ◽

Cell Lines ◽

Primary Cell Culture ◽

Tumor Tissue ◽

Primary Cell ◽

Glial Tumor ◽

Ki 67 ◽

Glial Tumors

e14515 Background: The choice of cell source for 3D bioprinting of in vivo-like models of glial tumors is crucial and must take into account the ability to proliferation and stable metabolism. Oral administration of 5-aminolevulinic acid (5-ALA) in patients prior to surgery increases the fluorescent contrast between tumor and surrounding tissue, but the effect of contrast agents on cells in vitro is unknown. The aim of the study was obtaining viable glial tumor tissues using 5-ALA, as well as the development of a stable primary cell culture for 3D bioprinting. Methods: Tumor tissue was obtained from patients with glioblastoma during surgery under visual control using the Opmi Pentero Blue E400 microscope and 5-ALA. Material was disaggregated on a BD Machine using Medicons 50 μm (BD). Glioblastoma cells were cultured in DMEM/F12 medium with L-glutamine (Gibco) containing 10% fetal bovine serum (Biolot, Russia), 1% non-essential amino acids (NEAA, Sigma-Aldrich) and 0.5% penicillin-streptomycin (Biolot) at 37C. Glial cell lines were characterized immunohistochemically using antibodies to the glial fibrillary acidic protein (GFAP) and proliferation index (Ki-67). Microsatellite analysis was performed using three dinucleotide repeat markers D2S123, D17S250, D5S346 and five mononucleotide loci BAT25, BAT26, NR21, NR24 and NR27. Results: The positive expression of GFAP on the cell processes of the star-like shape was clearly visualized, indicating a morphological feature of glial tumors. The Ki-67 labeling index was 70%. Changes were observed at the D17S250 locus (148-148/148-152) for the glial tumor primary cells after the sixth passage. Microsatellite instability was not observed in the primary cell culture. Conclusions: The accumulation of porphyrins from 5-ALA in glial tumor cells does not prevent the in vitro creation of a cell culture from tumor tissue. Microsatellite analysis showed that the obtained glioblastoma cell lines remain stable for at least 10 passages. Material obtained during resection using 5-ALA is a reliable source of stable glial tumor cell lines.

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