TMOD-01. CHARACTERIZATION OF PATIENT-DERIVED PRIMARY CELL LINES AND XENOGRAFTS FOR GLIOBLASTOMA

Abstract Patient-derived primary cell culture and xenograft are essential tools for translational research for glioblastoma. However, characteristics of each patient derived cell line and xenograft is not extensively studied. In this study, we aim to analyze the characteristics of our glioblastoma patient-derived cell lines and xenografts based on cell surface markers and their differentiation patterns. We have established 20 glioblastoma primary cell culture lines by serum free medium containing EGF and bFGF and found that primary cell culture lines could be classified based on the expression of CD133 and CD44. Four cell lines had high expression of both CD133 and CD44. Eleven cell lines had high expression of only CD44, three cell lines had high expression of only CD133, two cell lines had low expression of both CD133 and CD44. In addition when we induce differentiation, these cell lines showed differentiation to both glial and neuronal differentiation, but differentiation patterns were different depending on each cell line. Four cell lines showed predominant neuronal differentiation and others showed predominant glial differentiation. We next investigated in vivo characteristics of glioblastoma patient derived xenografts from these established cell lines. We have injected these cell lines into NOD/Shi-scid IL2Rγ KO mouse and histopathologically analyzed characteristics of xenografts. Each xenograft well recapitulated histological features of original patients’ tumors and tumor cells remarkably invade through subventricular zone. These results suggest that glioblastoma patient derived primary cell lines and xenografts have different characteristics of cell surface marker expressions and differentiation patterns, thus can classify these cell lines depending on cell surface marker expressions and differentiation patterns. Further analysis is needed to examine the biological importance of the differences in cell surface marker expressions and differentiation patterns.

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Development of Primary Cell Lines from Gill, Kidney, Spleen and Caudal Fin of Common Carp (Cyprinus carpio)

Jurnal Ilmiah Perikanan dan Kelautan ◽

10.20473/jipk.v12i1.17656 ◽

2020 ◽

Vol 12 (1) ◽

pp. 73

Author(s):

Insariani Insariani ◽

Trisniaty Trisniaty ◽

Freddy Riatmono ◽

Abdul Ghani

Keyword(s):

Cell Culture ◽

Cell Lines ◽

Common Carp ◽

Cyprinus Carpio ◽

Primary Cell ◽

Fish Cell ◽

Caudal Fin ◽

Common Carp Cyprinus Carpio ◽

Carp Cyprinus Carpio ◽

Primary Cell Lines

HighlihgtDevelop primary cultures derived From tissue tails fins, gills, kidney and spleen from local Indonesian carp (Cyprinus carpio).Primary culture cell with L15 Mediacell cultures consist of two type Fibroblast-like and epithelial –like cell AbstractThe fish cell lines technology have been developed for the interests of the fisheries world. This study aimed at developing a primary cell line from gill, kidney, spleen, and caudal fin of a common carp (Cyprinus carpio). A healthy common carp weighing 20 g (~1 month) was collected from the Cijeruk Fish Seed Center, Bogor. The development of primary cell lines from the gill, fin, tail, kidney and spleen tissue was performed in cell culture medium Leibovitz’s L-15 supplemented with 20% serum fetal bovine, 250 IU Penicillin, 250 µg / ml kanamycin sulfate and 2Mm L-Glutamine, and incubated at 28°C. Primary cell lines of caudal fin and gill began to form a monolayer on day 17 after culture. While the development of cell lines from kidney and spleen, although the initiation of cells and cells spread on the surface into a monolayer, was not perfect; therefore, the passage was unable to be done. Microscopic observations and Giemsa staining showed primary cell lines of caudal fin and gill based on cell morphology consisted of two cell types, fibroblast-like cells and epithelial-like cells. The first passage was done on day 17 when the confluence was more than 50%. The next passage was carried out every 3 weeks when confluence reached 70% -80%. The primary cell culture of gill was successfully passaged as much as 72 and the caudal fin was successfully passed as much as 89 times over 7 years. These new cell lines can be further used to propagate fish viruses and other biotechnology assays.

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Comparison of histological features seen in primary OSCC sites with three dimensional cell culture models of invasion using primary cell lines derived from the same patients

British Journal of Oral and Maxillofacial Surgery ◽

10.1016/j.bjoms.2012.04.228 ◽

2012 ◽

Vol 50 ◽

pp. S32

Author(s):

J. Dhanda ◽

J. Risk ◽

J. Stanbury ◽

J. Woolgar ◽

A. Triantafyllou ◽

...

Keyword(s):

Cell Culture ◽

Cell Lines ◽

Three Dimensional ◽

Primary Cell ◽

Histological Features ◽

Primary Cell Lines ◽

Culture Models ◽

Cell Culture Models ◽

Dimensional Cell

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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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A Method of Primary Cell Culture for Establishing Equine Long-Term Culture Cell Lines.

Journal of Equine Science ◽

10.1294/jes.8.95 ◽

1997 ◽

Vol 8 (4) ◽

pp. 95-99 ◽

Cited By ~ 3

Author(s):

Seiki WATANABE ◽

Youichirou ISHIKAWA ◽

Hiromi HARA ◽

Kei HANZAWA ◽

Harutaka MUKOYAMA

Keyword(s):

Cell Culture ◽

Cell Lines ◽

Primary Cell Culture ◽

Culture Cell ◽

Primary Cell ◽

Long Term Culture

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PRIMARY CELL CULTURE OF AEDES ALBOPICTUS MIDGUT CELLS: A PROSPECTIVE MODEL FOR IN VITRO STUDY OF ARBOVIRUSES

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i10.19136 ◽

2017 ◽

Vol 10 (10) ◽

pp. 223

Author(s):

Enakshi Roy ◽

Moonmoon Sinha ◽

Shailja Katoch ◽

Urmita Chakraborty ◽

Satadal Das ◽

...

Keyword(s):

Cell Culture ◽

Cell Line ◽

Cell Lines ◽

Primary Cell Culture ◽

In Vitro Study ◽

Primary Cell ◽

Human Beings ◽

Mosquito Midgut ◽

Columnar Cells

Objective: Midgut cells play a key role in the propagation of mosquito borne Arboviruses. The existing mosquito cell lines for studying viral pathogenesis are derived either from larvae or from eggs since there is no cell line available from the mosquito midgut. Therefore, to delineate the in situ viral interaction which naturally occurs within the mosquito midgut and represent cellular pathogenesis in human beings, the present work was aimed to develop a primary cell line from the midgut cells of Aedes albopictus.Methods: The midgut cells of A. albopictus were collected, cultured and incubated at 28°C to study the growth after every 24 hrs for 7 days.Result: The primary cell culture showed an increasing growth pattern of columnar cells up to 48 hrs followed by decrease in cell population afterward. However, the number of stem cells increased significantly throughout the study period, and their population outnumbered the columnar cells after 72 hrs. There was no significant change of goblet cells and regenerating cells which were scanty in number throughout the experiment.Conclusion: The present method will help to develop the individual cell lines from mosquito midgut and study the host pathogen interaction in arboviral diseases in future.

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Characterization of adherent primary cell lines from fresh human glioblastoma tissue, defining glial fibrillary acidic protein as a reliable marker in establishment of glioblastoma cell culture

Cancer Reports ◽

10.1002/cnr2.1324 ◽

2020 ◽

Author(s):

Susanne Grube ◽

Diana Freitag ◽

Rolf Kalff ◽

Christian Ewald ◽

Jan Walter

Keyword(s):

Cell Culture ◽

Glial Fibrillary Acidic Protein ◽

Cell Lines ◽

Acidic Protein ◽

Primary Cell ◽

Glioblastoma Cell ◽

Human Glioblastoma ◽

Primary Cell Lines ◽

Reliable Marker

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Preclinical evidence of ET-743 as a potential chemotherapy option for the treatment of biliary carcinoma.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.4_suppl.193 ◽

2013 ◽

Vol 31 (4_suppl) ◽

pp. 193-193

Author(s):

Francesco Leone ◽

Caterina Peraldo-Neia ◽

Giuliana Cavalloni ◽

Marco Soster ◽

Loretta Gammaitoni ◽

...

Keyword(s):

Cell Cycle ◽

Cell Culture ◽

Dna Damage ◽

Clinical Trials ◽

Cell Lines ◽

Primary Cell Culture ◽

Primary Cell ◽

Term Survival ◽

Anti Tumor Activity

193 Background: The standard chemotherapy for unresectablebiliary tract carcinoma (BTC) is based on gemcitabine and platinum compounds. However, these combinations have not been shown to be effective in improving long-term survival. Thus, there is a real need to find new strategies that would impact in a significant way on clinical outcome. Ecteinascidin-743 (ET-743), a compound isolated from the marine tunicate Ecteinascidia turbinata. ET-743, is approved for the treatment of ovarian cancer and soft tissue sarcoma. Phase II and III clinical trials are ongoing for the treatment of different solid tumors. No preclinical data are available about the efficacy of ET-743 in BTC. In a phase I study, one patient received ET-743 plus capecitabine and experienced a long lasting complete metabolic response. Here, we investigated the antitumor activity of ET-743 in preclinical BTC models. Methods: Four BTC cell lines TFK1, EGI-1, HuH28 and TGBC1 were used to evaluate the effect of ET-743 on proliferation, cell cycle, apoptosis and on the activation of DNA damage proteins. The effect on proliferation was also investigated on a primary cell culture of a gallbladder carcinoma (GBC) resistant to gemcitabine and oxaliplatin. On the same cells, the inhibition of VEGF secretion mediated by ET-743 was analyzed by ELISA. The anti-tumor activity of ET-743 was tested on EGI-1 xenografts in NOD/SCID mice. Results: In vitro, ET-743 is able to markedly reduce cell proliferation of BTC cell lines through cell cycle blockage on G0/G1 phase and to inhibit the growth of primary cell culture derived from GBC patient. Moreover, ET-743 promotes apoptosis by caspase 3 activation, activates proteins involved in DNA damage and reduces VEGF secretion. In the in vivo model, ET-743 is able to slow tumor growth in BTC xenograft. The mechanism of anti-tumor activity involves DNA damage, the induction of hypoxia transcription factor-1, and angiogenesis inhibition. ET-743 has no significant effect on apoptosis in vivo. Conclusions: These data suggest that ET-743 could represent an alternative chemotherapy for BTC treatment and encourage the development of clinical trials of ET-743 in BTC patients.

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