scholarly journals Long-Term Culture of Giardia lamblia in Cell Culture Medium Requires Intimate Association with Viable Mammalian Cells

2019 ◽  
Vol 87 (11) ◽  
Author(s):  
Theodore E. Nash
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Cell Lines ◽  
Giardia Lamblia ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Host Cells ◽  
Cell Culture Medium ◽  
Heterologous Cell

ABSTRACT Giardia lamblia is usually cultured axenically in TYI-S-33, a complex medium which does not permit survival and growth of mammalian cells. Likewise, medium commonly used to maintain and grow mammalian cells does not support healthy trophozoite survival for more than a few hours. The inability to coculture trophozoites and epithelial cells under optimal conditions limits studies of their interactions as well as interpretation of results. Trophozoites of the WB isolate but not the GS isolate were repeatedly adapted to grow stably in long-term cocultures with Caco2, Cos7, and mouse tumor rectal (RIT) cell lines using hybridoma-screened Dulbecco’s modified Eagle’s medium and 10% fetal calf serum. Giardia did not grow in spent cell culture medium or when separated by a permeable membrane using transwell methodology. Giardia chronically cocultured with specific cell lines became adapted (conditioned). These Giardia cocultures grew better than nonconditioned trophozoites, and the cell lines differed in their ability to support trophozoite growth in the order of RIT > Cos7 > Caco2. Trophozoites conditioned on one cell line and then grown in the presence of a heterologous cell line changed their growth rate to that seen in conditioned Giardia from the heterologous cell line. Trophozoite survival required intimate contact with cells, suggesting that trophozoites obtain an essential nutrient or growth factor from mammalian cells. This may explain why Giardia trophozoites adhere to the small intestinal epithelium during human and animal infections. This coculture system will be useful to understand the complex interactions between the host cells and parasite.

Adaptation of an insect cell line (Agallia constricta) in a mammalian cell culture medium

In Vitro ◽  
1973 ◽  
Vol 8 (5) ◽  
pp. 375-378 ◽  
Author(s):  
Arthur H. Intosh ◽  
K. Maramorosch ◽  
C. Rechtoris
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Mammalian Cell ◽  
Insect Cell ◽  
Culture Medium ◽  
Mammalian Cell Culture ◽  
Insect Cell Line ◽  
Cell Culture Medium

scholarly journals Analysis of a Novel Insect Cell Culture Medium-Based Growth Medium for Bartonella Species

2008 ◽  
Vol 74 (16) ◽  
pp. 5224-5227 ◽  
Author(s):  
Tanja Riess ◽  
Florian Dietrich ◽  
Katja V. Schmidt ◽  
Patrick O. Kaiser ◽  
Heinz Schwarz ◽  
...  
Keyword(s):  
Cell Culture ◽  
Insect Cell ◽  
Culture Medium ◽  
Expression Patterns ◽  
Insect Cell Culture ◽  
Host Cells ◽  
Cell Culture Medium ◽  
Bacterial Protein ◽  
Bartonella Species ◽  
Slow Growing

ABSTRACT Human- and animal-pathogenic Bartonella species are fastidious and slow-growing bacteria difficult to isolate and cultivate. We describe a novel, easy-to-prepare liquid medium for the fast and reliable growth of several Bartonella spp. that does not affect bacterial protein expression patterns or interactions with host cells.

scholarly journals The landscape of metabolic pathway dependencies in cancer cell lines

2021 ◽  
Vol 17 (4) ◽  
pp. e1008942
Author(s):  
James H. Joly ◽  
Brandon T. L. Chew ◽  
Nicholas A. Graham
Keyword(s):  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Metabolic Pathway ◽  
Culture Medium ◽  
Cancer Cell Lines ◽  
Metabolic Reprogramming ◽  
Cell Culture Medium ◽  
Loss Of Function

The metabolic reprogramming of cancer cells creates metabolic vulnerabilities that can be therapeutically targeted. However, our understanding of metabolic dependencies and the pathway crosstalk that creates these vulnerabilities in cancer cells remains incomplete. Here, by integrating gene expression data with genetic loss-of-function and pharmacological screening data from hundreds of cancer cell lines, we identified metabolic vulnerabilities at the level of pathways rather than individual genes. This approach revealed that metabolic pathway dependencies are highly context-specific such that cancer cells are vulnerable to inhibition of one metabolic pathway only when activity of another metabolic pathway is altered. Notably, we also found that the no single metabolic pathway was universally essential, suggesting that cancer cells are not invariably dependent on any metabolic pathway. In addition, we confirmed that cell culture medium is a major confounding factor for the analysis of metabolic pathway vulnerabilities. Nevertheless, we found robust associations between metabolic pathway activity and sensitivity to clinically approved drugs that were independent of cell culture medium. Lastly, we used parallel integration of pharmacological and genetic dependency data to confidently identify metabolic pathway vulnerabilities. Taken together, this study serves as a comprehensive characterization of the landscape of metabolic pathway vulnerabilities in cancer cell lines.

scholarly journals Addition of Exogenous Protease Facilitates Reovirus Infection in Many Restrictive Cells

2002 ◽  
Vol 76 (15) ◽  
pp. 7430-7443 ◽  
Author(s):  
Joseph W. Golden ◽  
Jessica Linke ◽  
Stephen Schmechel ◽  
Kara Thoemke ◽  
Leslie A. Schiff
Keyword(s):  
Cell Culture ◽  
Life Cycle ◽  
Host Range ◽  
Cell Lines ◽  
Culture Medium ◽  
Cathepsin L ◽  
Cell Culture Medium ◽  
Reovirus Infection ◽  
Lysosomal Protease ◽  
Mammalian Cell Lines

ABSTRACT Virion uncoating is a critical step in the life cycle of mammalian orthoreoviruses. In cell culture, and probably in extraintestinal tissues in vivo, reovirus virions undergo partial proteolysis within endosomal or/or lysosomal compartments. This process converts the virion into a form referred to as an intermediate subvirion particle (ISVP). In natural enteric reovirus infections, proteolytic uncoating takes place extracellularly within the intestinal lumen. The resultant proteolyzed particles, unlike intact virions, have the capacity to penetrate cell membranes and thereby gain access to cytoplasmic components required for viral gene expression. We hypothesized that the capacity of reovirus outer capsid proteins to be proteolyzed is a determinant of cellular host range. To investigate this hypothesis, we asked if the addition of protease to cell culture medium would expand the range of cultured mammalian cell lines that can be productively infected by reoviruses. We identified many transformed and nontransformed cell lines, as well as primary cells, that restrict viral infection. In several of these restrictive cells, virion uncoating is inefficient or blocked. Addition of proteases to the cell culture medium generates ISVP-like particles and promotes viral growth in nearly all cell lines tested. Interestingly, we found that some cell lines that restrict reovirus uncoating still express mature cathepsin L, a lysosomal protease required for virion disassembly in murine L929 cells. This finding suggests that factors in addition to cathepsin L are required for efficient intracellular proteolysis of reovirus virions. Our results demonstrate that virion uncoating is a critical determinant of reovirus cellular host range and that many cells which otherwise support productive reovirus infection cannot efficiently mediate this essential early step in the virus life cycle.

scholarly journals Adaptation of Cholesterol Requiring NS0 Cells to Serum Free Culture Conditions

2011 ◽  
Vol 12 (4) ◽  
Author(s):  
Junaid Muneer Raja ◽  
Nurina Anuar ◽  
And Badarulhisam Abdul Rahman
Keyword(s):  
Cell Culture ◽  
Monoclonal Antibodies ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Downstream Processing ◽  
Western World ◽  
Human Consumption ◽  
Cell Culture Medium ◽  
Serum Free ◽  
Ns0 Cells

Colorectal cancer is the third most common form of cancer and the second leading cause of cancer-related death in the Western world. The answers to such life threatening diseases and cancers are monoclonal antibodies (MAb's) which are widely used as therapeutic agents. World demand for currently approved MAb's is on the order of a few kilograms per year. However, new therapeutic MAb's are under development and require doses of several hundred milligrams to a gram over the course of therapy. Very often to cater for the special requirements for the growth of mammalian cells, serum is added to the cell culture medium. However, removal of serum from the cell culture medium is often carried out, especially if the end product is to be used for human consumption, in order to eliminate various disadvantages such as high physiological variability, high batch to batch variability, risk of contamination and high cost, and challenges posed in the downstream processing of the product. In this paper, the adaptation of cholesterol requiring NS0 cells to commercially available serum free media is presented. ABSTRAK: Kanser kolorektum merupakan kanser ketiga paling umum dan kini berada di tempat kedua penyebab kematian berkaitan kanser di negara Barat. Jawapan kepada penyakit yang mengancam nyawa dan penyakit kanser adalah antibodi monoklon (monoclonal antibodies ((MAb's)) yang digunakan sebagai agen terapeutik. Permintaan dunia terhadap MAb's yang diluluskan adalah dalam bilangan beberapa kilogram setahun. Namun, terapeutik MAb's yang baru adalah di bawah penyelidikan dan memerlukan beberapa ratus dos milligram hingga satu gram dalam satu peringkat terapi. Sering kali untuk memenuhi permintaan terhadap tumbesaran sel mamalia, serum dicampurkan dengan sel kultur perantara. Walaupun begitu, pemindahan serum dari sel kultur perantara sering dilakukan, terutamanya jika produk akhir digunakan untuk kegunaan manusia; untuk mengurangkan pelbagai kelemahan seperti kebolehubahan psikologi yang tinggi, kebolehubahan yang tinggi daripada satu kumpulan ke satu kumpulan lain, risiko pencemaran, kos yang tinggi, dan cabaran mendatang dalam pemprosesan produk. Dalam perbentangan ini, kolestrol yang diubah memerlukan sel NS0 yang dikomersilkan dengan serum bebas perantara.

scholarly journals The landscape of metabolic pathway dependencies in cancer cell lines

2020 ◽  
Author(s):  
James H. Joly ◽  
Brandon T.L. Chew ◽  
Nicholas A. Graham
Keyword(s):  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Metabolic Pathway ◽  
Culture Medium ◽  
Cancer Cell Lines ◽  
Metabolic Reprogramming ◽  
Cell Culture Medium ◽  
Loss Of Function

AbstractThe metabolic reprogramming of cancer cells creates metabolic vulnerabilities that can be therapeutically targeted. However, our understanding of metabolic dependencies and the pathway crosstalk that creates these vulnerabilities in cancer cells remains incomplete. Here, by integrating gene expression data with genetic loss-of-function and pharmacological screening data from hundreds of cancer cell lines, we identified metabolic vulnerabilities at the level of pathways rather than individual genes. This approach revealed that metabolic pathway dependencies are highly context-specific such cancer cells are vulnerable to inhibition of one metabolic pathway only when activity of another metabolic pathway is altered. Notably, we also found that the no single metabolic pathway was universally essential, suggesting that cancer cells are not invariably dependent on any metabolic pathway. In addition, we confirmed that cell culture medium is a major confounding factor for the analysis of metabolic pathway vulnerabilities. Nevertheless, we found robust associations between metabolic pathway activity and sensitivity to clinically approved drugs that were independent of cell culture medium. Lastly, we used parallel integration of pharmacological and genetic dependency data to confidently identify metabolic pathway vulnerabilities. Taken together, this study serves as a comprehensive characterization of the landscape of metabolic pathway vulnerabilities in cancer cell lines.

GTSF-2: A new, versatile cell culture medium for diverse normal and transformed mammalian cells

1997 ◽  
Vol 33 (5) ◽  
pp. 344-351 ◽  
Author(s):  
Peter I. Lelkes ◽  
Esther Ramos ◽  
Victor V. Nikolaychik ◽  
Dawn M. Wankowski ◽  
Brian R. Unsworth ◽  
...  
Keyword(s):  
Cell Culture ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Cell Culture Medium

The effect of four lanthanides onto a rat kidney cell line (NRK-52E) is dependent on the composition of the cell culture medium

Toxicology ◽  
2021 ◽  
Vol 456 ◽  
pp. 152771
Author(s):  
Anne Heller ◽  
Alina Pisarevskaja ◽  
Nora Bölicke ◽  
Astrid Barkleit ◽  
Frank Bok ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Kidney Cell ◽  
Culture Medium ◽  
Rat Kidney ◽  
Cell Culture Medium ◽  
Kidney Cell Line

Cytotoxicity of Metal Cations used in Dental Cast Alloys

1997 ◽  
Vol 25 (3) ◽  
pp. 323-330 ◽  
Author(s):  
Gottfried Schmalz ◽  
Dorthe Arenholt-Bindslev ◽  
Silke Pfüller ◽  
Helmut Schweikl
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Culture Medium ◽  
Rank Order ◽  
Metal Cations ◽  
Human Gingival Fibroblasts ◽  
Cell Culture Medium ◽  
Gingival Fibroblasts ◽  
Dental Alloys ◽  
Continuous Cell Lines

The toxicity of dental alloys depends upon the quantity and chemical state of metal elements released from the alloy into the affected tissue. The purpose of the present study was to investigate the effects of various cell lines, various cell numbers, and various serum concentrations on the cytotoxicity of metal cations which are components of dental alloys. Aqueous salt solutions were tested on two continuous cell lines (L-929 mouse fibroblast-like cells and HaK hamster epithelial-like kidney cells) and on primary human gingival fibroblasts. The cell culture medium was routinely supplemented with 5% fetal calf serum. The influence of the serum concentration on the cytotoxic potencies of metal cations was investigated by increasing the concentration to 20% in one series of experiments with L-929 cells. Cell reactions were recorded with the MTT test after exposure for 24 hours. The rank order of the cytotoxicity of cations in L-929 cells (10,000 cells/well, 5% serum) from the most to the least toxic was: Ag+, Zn2+, Cd2+, Hg2+, Au3+, Pt4+, Co2+, Cu2+, Ni2+, Pd2+, Mn2+, Nb5+, Mo5+, Ga3+, Cr3+, In3+, Sn2+. A variation in cell number between 5000 and 10,000 cells/well did not influence the results. Furthermore, no differences were observed in the cytotoxic responses of the two continuous cell lines. Variation in the serum content of L-929 cell culture medium had no significant influence on the cytotoxicity of most metal cations, except for Nb5+, Ni2+, Sn2+. Primary human gingival fibroblasts showed the same rank order of toxicity as the continuous cell lines, but at higher concentrations of the test materials.

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