INSECT CHROMOSOME ISOLATION: A METHOD FOR METAPHASE CHROMOSOME HARVEST FROM CULTURED CELLS OF THE MOSQUITO AEDES ALBOPICTUS

1981 ◽  
Vol 23 (3) ◽  
pp. 453-457 ◽  
Author(s):  
Asit B. Mukherjee ◽  
Lorraine DeGiorgio
Keyword(s):  
Cell Line ◽  
Aedes Albopictus ◽  
Metaphase Chromosome ◽  
Cultured Cells ◽  
Modified Technique ◽  
Metaphase Chromosomes ◽  
Chromosome Isolation

A modified technique for bulk isolation of metaphase chromosomes from an in vitro cell line of the mosquito Aedes albopictus (Skuse) is described.

scholarly journals Characterization of a Chinese Hamster Ovary Cell Line Developed by Retroviral Insertional Mutagenesis That Is Resistant to Sindbis Virus Infection

1999 ◽  
Vol 73 (6) ◽  
pp. 4919-4924 ◽  
Author(s):  
Jia-Tsrong Jan ◽  
Andrew P. Byrnes ◽  
Diane E. Griffin
Keyword(s):  
Heparan Sulfate ◽  
Cell Line ◽  
Virus Replication ◽  
Chinese Hamster Ovary ◽  
Sindbis Virus ◽  
Insertional Mutagenesis ◽  
Cultured Cells ◽  
Chinese Hamster ◽  
Ovary Cell

ABSTRACT The alphavirus Sindbis virus (SV) has a wide host range and infects many types of cultured cells in vitro. The outcome of infection is dependent on the strain of virus used for infection and the properties of the cells infected. To identify cellular determinants of susceptibility to SV infection we mutagenized Chinese hamster ovary (CHO) cells by retroviral insertion with a vector containing the neomycin resistance gene that allowed selection for integration into transcriptionally active genes. Cells were then selected for survival after infection with SV. The most resistant cell line (CHO-18.4m) exhibited delayed virus replication and virus-induced cell death, had a single retroviral insertion, and was defective in SV binding to the cell surface. Further analysis revealed that CHO-18.4m cells were deficient in the expression of the sulfated glycosaminoglycans heparan sulfate and chondroitin sulfate. This further confirms the importance of heparan sulfate as an attachment molecule for SV in vitro and demonstrates the usefulness of this technique for identifying cellular genes that are important for virus replication.

Infection, growth and maintenance of Wolbachia pipientis in clonal and non-clonal Aedes albopictus cell cultures

2012 ◽  
Vol 103 (3) ◽  
pp. 251-260 ◽  
Author(s):  
C.C.H. Khoo ◽  
C.M.P. Venard ◽  
Y. Fu ◽  
D.R. Mercer ◽  
S.L. Dobson
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Aedes Albopictus ◽  
Cell Types ◽  
Infection Level ◽  
Wolbachia Pipientis ◽  
Albopictus Cell ◽  
Host Cell Interactions ◽  
Insect Cell Lines

AbstractInsect cell lines provide useful in vitro models for studying biological systems, including interactions between mosquitoes and obligate intracellular endosymbionts such as Wolbachia pipientis. The Aedes albopictus Aa23 cell line was the first cell line developed to allow examination of Wolbachia infections. However, Wolbachia studies using Aa23 can be complicated by the presence of different cell types in the cell line and the substantial temporal variation in infection level. Two approaches were examined to ameliorate infection variability. In the first approach, multiple Aa23 passaging regimes were tested for an effect on infection variability. Fluorescence in situ hybridization (FISH) staining was used to characterize Wolbachia infection level over time. The results demonstrate an impact of passaging method on Wolbachia infection level, with some methods resulting in loss of infection. None of the passaging methods succeeded in effectively mitigating infection level variation. In a second approach, the clonal C7-10 A. albopictus cell line was infected with Wolbachia from Aa23 cells and Drosophila simulans (Riverside), resulting in cell lines designated C7-10B and C7-10R, respectively. Characterization via FISH staining showed greater stability and uniformity of Wolbachia infection in C7-10R relative to the infection in C7-10B. Characterization of the Aa23, C7-10B and C7-10R lines is discussed as a tool for the study of Wolbachia-host cell interactions.

Immunophenotyping of Nonneoplastic and Neoplastic Histiocytes in Cats and Characterization of a Novel Cell Line Derived From Feline Progressive Histiocytosis

2020 ◽  
Vol 57 (6) ◽  
pp. 758-773
Author(s):  
Miyuki Hirabayashi ◽  
James K. Chambers ◽  
Ayumi Sumi ◽  
Kei Harada ◽  
Makoto Haritani ◽  
...  
Keyword(s):  
Cell Line ◽  
Cultured Cells ◽  
Survival Times ◽  
Neoplastic Cells ◽  
Hla Dr ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
E Cadherin

Histiocytic proliferative diseases are rare in cats, and their pathogenesis is poorly understood. In the present study, 25 cases of histiocytic sarcoma (HS) and 6 of feline progressive histiocytosis (FPH) were examined, and survival times were recorded in 19 cases. The immunophenotypes of tumor cells in these cases as well as of nonneoplastic feline histiocytes were characterized using formalin-fixed, paraffin-embedded tissues. An FPH cell line (AS-FPH01) and xenotransplant mouse model of FPH were also established. The median survival time of HS (150 days) was significantly shorter than that of FPH (470 days). Immunohistochemically, nonneoplastic histiocytes were immunopositive for various combinations of Iba-1, HLA-DR, E-cadherin, CD204, CD163, CD208, and MAC387. By immunohistochemistry, dermal interstitial dendritic cells (iDCs) and macrophages were CD204+/E-cadherin−, while epidermal Langerhans cells (LCs) were CD204−/E-cadherin+. Neoplastic cells of 4 FPH and 18 HS were CD204+/E-cadherin− (iDC/macrophage immunophenotype), while 2 FPH and 2 HS were CD204−/E-cadherin+ (LC immunophenotype), and 5 HS were CD204+/E-cadherin+ (LC-like cell immunophenotype). Furthermore, immunohistochemical and western blot analyses of AS-FPH01 cells derived from E-cadherin-negative FPH revealed that cultured cells were immunopositive for both CD204 and E-cadherin in vitro and in vivo. These results indicate that the neoplastic cells of feline HS and FPH were variably positive for iDC/macrophage and LC markers, and their immunophenotype changed in different microenvironments. The novel cell line established in the present study may serve as an experimental model of FPH that will enable further molecular and therapeutic studies on this disease.

scholarly journals Establishment and characterization of a new human eosinophilic leukemia cell line

Blood ◽  
1985 ◽  
Vol 66 (6) ◽  
pp. 1233-1240 ◽  
Author(s):  
H Saito ◽  
A Bourinbaiar ◽  
M Ginsburg ◽  
K Minato ◽  
E Ceresi ◽  
...  
Keyword(s):  
Cell Line ◽  
Cultured Cells ◽  
Interleukin 2 ◽  
Philadelphia Chromosome ◽  
Leukemia Cell ◽  
Culture Conditions ◽  
Membrane Receptors ◽  
Leukemia Cell Line ◽  
Standard Culture

Abstract A human eosinophilic leukemia cell line, designated as EoL, was established from the peripheral blood of a patient with Philadelphia chromosome-negative eosinophilic leukemia (EL). The EoL cell line grows in single cell suspension with a doubling time of 48 hours for about one year. The reactivity of these cells was tested with a panel of monoclonal antibodies; they were found to express surface IA antigen, myeloid antigen (IF10, MY9) and membrane receptors for interleukin 2 (IL-2, Tac antigen). Under standard culture conditions, a small percentage of cells having more typical eosinophilic characteristics was present. These cells had cytoplasmic granules and were positive for Luxol-fast-blue and eosinophil peroxidase. Under culture conditions to induce the maturation of myeloid cells, such as alkaline medium or addition of dimethyl sulfoxide (DMSO), the frequency of cells with typical eosinophilic features increased to about 40%. In addition, cytogenetic studies showed that cultured cells and original leukemic blasts presented similar chromosome abnormalities. EoL seems to be a unique leukemic line committed to the eosinophilic lineage and can provide a useful in vitro model for the study of malignant eosinophilic properties.

In vitro propagation of hepatopancreatic parvo-like virus (HPV) of shrimp in C6/36 (Aedes albopictus) cell line

2013 ◽  
Vol 112 (3) ◽  
pp. 229-235 ◽  
Author(s):  
N. Madan ◽  
K.S.N. Nambi ◽  
S. Abdul Majeed ◽  
G. Taju ◽  
N. Sundar Raj ◽  
...  
Keyword(s):  
Cell Line ◽  
Aedes Albopictus ◽  
In Vitro Propagation ◽  
Albopictus Cell

scholarly journals Analysis of the Aedes albopictus C6/36 genome provides insight into cell line adaptations to in vitro viral propagation

2017 ◽  
Author(s):  
Jason R Miller ◽  
Sergey Koren ◽  
Kari A Dilley ◽  
Vinita Puri ◽  
David M Brown ◽  
...  
Keyword(s):  
Cell Line ◽  
Aedes Albopictus ◽  
Sequence Data ◽  
Gene Annotation ◽  
Virus Transmission ◽  
Data Set ◽  
Rna Sequence ◽  
Viral Propagation ◽  
Male Specific

ABSTRACTBackgroundThe 50-year old Aedes albopictus C6/36 cell line is a resource for the detection, amplification, and analysis of mosquito-borne viruses including Zika, dengue, and chikungunya. The cell line is derived from an unknown number of larvae from an unspecified strain of Aedes albopictus mosquitoes. Toward improved utility of the cell line for research in virus transmission, we present an annotated assembly of the C6/36 genome.ResultsThe C6/36 genome assembly has the largest contig N50 (3.3 Mbp) of any mosquito assembly, presents the sequences of both haplotypes for most of the diploid genome, reveals independent null mutations in both alleles of the Dicer locus, and indicates a male-specific genome. Gene annotation was computed with publicly available mosquito transcript sequences. Gene expression data from cell line RNA sequence identified enrichment of growth-related pathways and conspicuous deficiency in aquaporins and inward rectifier K+ channels. As a test of utility, RNA sequence data from Zika-infected cells was mapped to the C6/36 genome and transcriptome assemblies. Host subtraction reduced the data set by 89%, enabling faster characterization of non-host reads.ConclusionsThe C6/36 genome sequence and annotation should enable additional uses of the cell line to study arbovirus vector interactions and interventions aimed at restricting the spread of human disease.

scholarly journals Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Behnaz Ashtari ◽  
Azar Shams ◽  
Narges Esmaeilzadeh ◽  
Sara Tanbakooei ◽  
Morteza Koruji ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Line ◽  
Microfluidic Device ◽  
Cultured Cells ◽  
Initial Step ◽  
Spermatogonial Stem Cells ◽  
Control Group ◽  
Isolation And Purification ◽  
Laboratory Technique

Abstract Background Some children who have survived cancer will be azoospermic in the future. Performing isolation and purification procedures for spermatogonial stem cells (SSC) is very critical. In this regard, performing the process of decontamination of cancerous cells is the initial step. The major objective of the present study is to separate the malignant EL4 cell line in mice and spermatogonial stem cells in vitro. Methods The spermatogonial stem cells of sixty neonatal mice were isolated, and the procedure of co-culturing was carried out by EL4 which were classified into 2 major groups: (1) the control group (co-culture in a growth medium) and (2) the group of co-cultured cells which were separated using the microfluidic device. The percentage of cells was assessed using flow cytometry technique and common laboratory technique of immunocytochemistry and finally was confirmed through the laboratory technique of reverse transcription-polymerase chain reaction (RT-PCR). Results The actual percentage of EL4 and SSC after isolation was collected at two outlets: the outputs for the smaller outlet were 0.12% for SSC and 42.14% for EL4, while in the larger outlet, the outputs were 80.38% for SSC and 0.32% for EL4; in the control group, the percentages of cells were 21.44% for SSC and 23.28% for EL4 (based on t test (p ≤ 0.05)). Conclusions The present study demonstrates that the use of the microfluidic device is effective in separating cancer cells from spermatogonial stem cells.

In vitro cultivation of Wolbachia pipientis in an Aedes albopictus cell line

1997 ◽  
Vol 6 (1) ◽  
pp. 33-39 ◽  
Author(s):  
S. L. O'Neill ◽  
M. M. Pettigrew ◽  
S. P. Sinkins ◽  
H. R. Braig ◽  
T. G. Andreadis ◽  
...  
Keyword(s):  
Cell Line ◽  
Aedes Albopictus ◽  
In Vitro Cultivation ◽  
Wolbachia Pipientis ◽  
Albopictus Cell

INSECT CHROMOSOME BANDING: TECHNIQUE FOR G- AND Q-BANDING PATTERN IN THE MOSQUITO AEDES ALBOPICTUS

1975 ◽  
Vol 17 (2) ◽  
pp. 241-244 ◽  
Author(s):  
Gerald E. Steiniger ◽  
Asit B. Mukherjee
Keyword(s):  
Aedes Albopictus ◽  
Banding Pattern ◽  
Chromosome Banding ◽  
Banding Technique ◽  
Modified Technique ◽  
Metaphase Chromosomes ◽  
Somatic Metaphase

A modified technique is described for the production of clear G- and Q-bands of somatic metaphase chromosomes of the mosquito, Aedes albopictus Skuse.

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