Ultrastructural and immunocytochemical characterization of cultured cells from rat molar stellate reticulum

1990 ◽  
Vol 35 (8) ◽  
pp. 603-613 ◽  
Author(s):  
G.E. Wise ◽  
V.L. Rudick ◽  
A.M. Brun-Zinkernagel ◽  
W. Fan
Keyword(s):  
Cultured Cells ◽  
Stellate Reticulum ◽  
Rat Molar

scholarly journals Growth, lifetime, directional movement and myosin-dependent motility of mutant keratin granules in cultured cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
S. M. Lehmann ◽  
R. E. Leube ◽  
R. Windoffer
Keyword(s):  
Digital Image Analysis ◽  
Cultured Cells ◽  
Epidermolysis Bullosa Simplex ◽  
Continuous Growth ◽  
Relative Contribution ◽  
Complete Life Cycle ◽  
Directional Movement ◽  
Multiple Characteristics ◽  
Cell Centre

AbstractIntermediate filament polypeptides (IFPs) are prominent components of cytoplasmic aggregates, which are pathognomonic for multiple diseases. Recent observations in cultured cells suggest that they are dynamic and subject to regulated turnover. The emerging concept is that multiple factors contribute to motility and turnover of IFP-containing aggregates. To understand their relative contribution, quantitative tools are needed. The current study addresses this need using epithelial cells producing mutant keratin IFPs that have been identified as the cause of the hereditary blister-forming skin disease epidermolysis bullosa simplex. Digital image analysis of individual granules allowed mapping of their complete life cycle, with information on multiple characteristics at any given time-point. The deduced signet features revealed rapid granule fusion and directed transport from the periphery towards the cell centre, and a limited, ~ 30 min lifetime with a slow, continuous growth phase followed by fast disassembly. As paradigmatic proof-of-principle, we demonstrate that inhibition of myosin II selectively reduces granule movement, linking keratin granule motility to retrograde cortical acto-myosin flow. The newly developed methods and established parameters will help in the characterization of known and the identification of novel regulators of IFP-containing aggregates.

scholarly journals Immortalization and Characterization of Rat Lingual Keratinocytes in a High-Calcium and Feeder-Free Culture System Using ROCK Inhibitor Y-27632

2021 ◽  
Vol 22 (13) ◽  
pp. 6782
Author(s):  
Zixing Chen ◽  
Wenmeng He ◽  
Thomas Chun Ning Leung ◽  
Hau Yin Chung
Keyword(s):  
Calcium Imaging ◽  
Cultured Cells ◽  
Rho Kinase ◽  
Culture Method ◽  
Taste Bud ◽  
Label Free ◽  
Sprague Dawley ◽  
Rock Inhibitor ◽  
High Calcium

Cultured keratinocytes are desirable models for biological and medical studies. However, primary keratinocytes are difficult to maintain, and there has been little research on lingual keratinocyte culture. Here, we investigated the effect of Y-27632, a Rho kinase (ROCK) inhibitor, on the immortalization and characterization of cultured rat lingual keratinocyte (RLKs). Three Y-27632–supplemented media were screened for the cultivation of RLKs isolated from Sprague–Dawley rats. Phalloidin staining and TUNEL assay were applied to visualize cytoskeleton dynamics and cell apoptosis following Y-27632 removal. Label-free proteomics, RT-PCR, calcium imaging, and cytogenetic studies were conducted to characterize the cultured cells. Results showed that RLKs could be conditionally immortalized in a high-calcium medium in the absence of feeder cells, although they did not exhibit normal karyotypes. The removal of Y-27632 from the culture medium led to reversible cytoskeletal reorganization and nuclear enlargement without triggering apoptosis, and a total of 239 differentially expressed proteins were identified by proteomic analysis. Notably, RLKs derived from the non-taste epithelium expressed some molecular markers characteristic of taste bud cells, yet calcium imaging revealed that they rarely responded to tastants. Collectively, we established a high-calcium and feeder-free culture method for the long-term maintenance of RLKs. Our results shed some new light on the immortalization and differentiation of lingual keratinocytes.

Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum

Cell ◽  
1978 ◽  
Vol 13 (2) ◽  
pp. 249-261 ◽  
Author(s):  
William E. Gordon ◽  
Anne Bushnell ◽  
Keith Burridge
Keyword(s):  
Cultured Cells ◽  
Rabbit Antiserum ◽  
10 Nm Filaments

scholarly journals Establishment and characterization of fibroblast cultures derived from a female common hippopotamus (Hippopotamus amphibius) skin biopsy

2020 ◽  
Author(s):  
Tao Wang ◽  
Zelong Li ◽  
Jinpu Wei ◽  
Dongmin Zheng ◽  
Chen Wang ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Cultured Cells ◽  
Population Decline ◽  
Fibroblast Cell ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Fibroblast Cultures ◽  
Hippopotamus Amphibius ◽  
The Common

AbstractThe population decline in the common hippopotamus (Hippopotamus amphibius) has necessitated the preservation of their genetic resources for species conservation and research. Of all actions, cryopreservation of fibroblast cell cultures derived from animal biopsy is considered a simple but efficient means. Nevertheless, preserving viable cell cultures of the common hippopotamus has not been achieved to our knowledge. To this end, we detailed a method to establish fibroblast cell cultures from a female common hippopotamus fetus in this study. By combining the classic tissue explant direct culture and enzymatic digestion methods, we isolated a great number of cells with typical fibroblastic morphology and high viability. Characterization of the fibroblast cultures was carried out using different techniques. In short, neither bacteria/fungi nor mycoplasma was detectable in the cell cultures throughout the study. The population doubling time was 23.9 h according to the growth curve. Karyotyping based on Giemsa staining showed that cultured cells were diploid with 36 chromosomes in all, one pair of which was sex chromosomes. Mitochondrial cytochrome C oxidase subunit I gene sequence of the cultured cells was 99.26% identical with the Hippopotamus amphibius complete mitochondrial DNA sequence registered in GenBank, confirming the cells were derived from a common hippopotamus. Flow cytometry and immunofluorescence staining results revealed that the detected cells were positive for fibroblast markers, S100A4 and Vimentin. In conclusion, we isolated and characterized a new fibroblast cell culture from a common hippopotamus skin sample and the cryopreserved cells could be useful genetic materials for the future research.

Isolation and Characterization of UDP-glucosyltransferase in the Cultured Cells of Nicotiana tabacum

Heterocycles ◽  
2002 ◽  
Vol 56 (1-2) ◽  
pp. 509
Author(s):  
Toshifumi Hirata ◽  
Shin-ya Yamane ◽  
Kei Shimoda ◽  
Takeshi Fujino ◽  
Shinji Ohta
Keyword(s):  
Nicotiana Tabacum ◽  
Cultured Cells ◽  
Isolation And Characterization

scholarly journals Generation of Glioblastoma Patient-Derived Intracranial Xenografts for Preclinical Studies

2020 ◽  
Vol 21 (14) ◽  
pp. 5113
Author(s):  
Amber E. Kerstetter-Fogle ◽  
Peggy L. R. Harris ◽  
Susann M. Brady-Kalnay ◽  
Andrew E. Sloan
Keyword(s):  
Cultured Cells ◽  
Clonal Selection ◽  
Previous Treatment ◽  
Growth Patterns ◽  
Phenotypic Characterization ◽  
Genomic Signatures ◽  
Culture Model ◽  
Passage Number ◽  
Human Gbm

Glioblastoma multiforme (GBM) is the most malignant primary brain cancer affecting adults. Therapeutic options for GBM have remained the same for over a decade with no significant improvement. Many therapies that are successful in culture have failed in patients, likely due to the complex microenvironment in the brain, which has yet to be reproduced in any culture model. Furthermore, the high passage number of cultured cells and clonal selection fail to recapitulate the molecular and genomic signatures of GBM. We have established orthotopic patient-derived xenografts (PDX) from 37 GBM patients with human GBM. Of the 69 patient samples analyzed, we were successful in passaging 37 lines three or more generations (53.6%). After phenotypic characterization of the xenografted tumor tissue, two different growth patterns emerged highly invasive or localized. The phenotype was dependent on malignancy and previous treatment of the patient from which the xenograft was derived. Physiologically, mice exhibited symptoms more quickly with each subsequent passage, particularly in the localized tumors. Study of these physiologically relevant human xenografts in mice will enable therapeutic screenings in a microenvironment that more closely resembles GBM and may allow development of individualized patient models which may eventually be used for simulating treatment.

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