Growth characteristics of a cell line derived from the pig oviduct

1975 ◽  
Vol 21 (12) ◽  
pp. 2094-2097 ◽  
Author(s):  
A. M. P. Bouillant ◽  
P. Genest ◽  
A. S. Greig
Keyword(s):  
Cell Line ◽  
Optimal Growth ◽  
Growth Conditions ◽  
Growth Characteristics ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Temperature Growth ◽  
A Cell ◽  
Maximum Population Density ◽  
Optimal Ph

Further evidence for the establishment of the pig fallopian tube (PFT) cell line as a continuous cell line was shown by an increase in the maximum population density as the number of subcultures increased. The optimal pH and temperature–growth ranges appeared to be 7.4–7.8 and 37–41 °C respectively, and the population doubling time was 20–25 h under optimal growth conditions. With progressive subculture, the serum requirements dropped from 20 to 2%. A plating efficiency of 2 to 4% was found in all serial subcultures. Colonies were observed in agar suspension culture at the 146th subculture and thereafter. Chromosomal alterations were found in the 100th subculture and thereafter.

Characterization of a newly established uterine carcinosarcoma cell line featuring the sarcomatous phenotype of the tumor in vitro

2008 ◽  
Vol 18 (2) ◽  
pp. 339-344 ◽  
Author(s):  
H.-J. Schulten ◽  
J. Wolf-Salgó ◽  
C. Gründker ◽  
B. Gunawan ◽  
L. FÜZESI
Keyword(s):  
Cell Line ◽  
Hormone Receptors ◽  
Tumor Biology ◽  
Growth Factor Receptor ◽  
Population Doubling ◽  
Established Cell Line ◽  
Population Doubling Time ◽  
Uterine Carcinosarcoma ◽  
Specific Staining

We describe the newly established cell line CS-99 derived from a uterine carcinosarcoma retaining features of the sarcomatous phenotype in vitro. CS-99 cells exhibit a mesenchymal morphology with predominantly spindle-shaped cells at nonconfluence turning to pleomorphic appearance at confluence. The mesenchymal phenotype was evidenced immunohistochemically by strong vimentin and moderate SM-actin, which was similar to the sarcomatous component of the primary tumor. P53 was overexpressed in a subset of CS-99 cells. Epithelial membrane antigen was moderately expressed whereas other markers including pan CK, CK 5/6, CK 34, epidermal growth factor receptor, desmin, carcinoembryonic antigen, S100, KIT, ERBB2, and the hormone receptors, estrogen receptor and progesterone receptor revealed either weak or no specific staining in CS-99 cells. High self-renewal capacity corresponded to the population doubling time of 23 h in high passage. CS-99 cells were able to develop three-dimensional tumor spheroids in vitro. Cytogenetic analysis and multicolor fluorescence in situ hybridization of CS-99 demonstrated an almost stable karyotype including numerical changes +8, +18, and +20 and translocations, amongst others der(1)t(1;2), der(1)t(1;7), der(2)t(2;19), der(5)t(5;8), and der(5)t(5;14). Taken together, the cell line CS-99 exhibits strong growths dynamics and a complex but stable karyotype in higher passages, and can be further a useful in vitro model system for studying tumor biology of carcinosarcomas.

scholarly journals Low Oxygen Tension Potentiates Proliferation and Stemness but Not Multilineage Differentiation of Caprine Male Germline Stem Cells

2021 ◽  
Author(s):  
Shiva Pratap Singh ◽  
Suresh Dinkar Kharche ◽  
Manisha Pathak ◽  
Ravi Ranjan ◽  
Yogesh Kumar Soni ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ex Vivo ◽  
Growth Characteristics ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Germline Stem Cells ◽  
Multilineage Differentiation ◽  
Low Oxygen ◽  
Differentiation Potential

Abstract The milieu of testicular germline stem cells (mGSCs) is characterized as low oxygen (O2) environment, whereas, there in-vitro expansion is typically performed under normoxia (20-21% O2). Here, we evaluated and compared the culture and multilineage differentiation characteristics of enriched (through differential platting and percoll density centrifugation) caprine mGSCs (cmGSCs) under hypoxic (5% O2) and normoxic (21% O2) culture conditions. For this, in addition to growth characteristics and population-doubling time (PDT); viability, proliferation, senescence, and expression of key-markers of adhesion (β-integrin and E-Cadherin) and stemness (OCT-4, THY-1 and UCHL-1) were evaluated and compared under normoxia and hypoxia. Moreover, the extent of multilineage differentiation (neurogenic, adipogenic, and chondrogenic differentiation) was assessed. The survival, viability and proliferation were significantly promoted and PDT was reduced (p < 0.05), thus yielding a higher number of viable cells with larger colonies under hypoxia. Furthermore, expression of stemness and adhesion markers was distinctly increased under lowered O2 condition. Conversely, the presence of differentiated regions and expression of differentiation specific key genes [C/EBPα (adipogenic), nestin and β-tubulin (neurogenic), and COL2A1 (chondrogenic)] were significantly (p < 0.05) reduced under hypoxic conditions. These data demonstrate that culturing cmGSCs under hypoxia augments the growth characteristics, and stemness but not the multilineage differentiation potential of cmGSCs as compared with normoxia. These data are important for the development of robust methodologies for ex-vivo expansion and lineage-committed differentiation of cmGSCs for clinical applications.

scholarly journals Developing an Ephestia kuehniella Hemocyte Cell Line to Assess the Bio-Insecticidal Potential of Microencapsulated Helicoverpa armigera Nucleopolyhedrovirus Against Cotton Bollworm (Lepidoptera: Noctuidae) Larva

2020 ◽  
Vol 113 (5) ◽  
pp. 2086-2095
Author(s):  
Bita Valizadeh ◽  
Samira Samarfard ◽  
Jalal Jalali Sendi ◽  
Thomas P Karbanowicz
Keyword(s):  
Cell Line ◽  
Helicoverpa Armigera ◽  
Ephestia Kuehniella ◽  
Cotton Bollworm ◽  
Population Doubling ◽  
Logarithmic Phase ◽  
Population Doubling Time ◽  
Cottonseed Kernel ◽  
Helicoverpa Armígera ◽  
Lepidoptera Noctuidae

Abstract Helicoverpa armigera Nucleopolyhedrovirus (HearNPV) (genus: Alphabaculovirus, incertae sedis: Baculoviridae) has been used to control Helicoverpa armigera (Hübner). A reproducible and susceptible cell line was prepared from the hemocytes of Ephestia kuehniella in Grace and Ex-Cell 420 media. The population doubling time of these cloned cell cultures during the logarithmic phase were about 2.3 and 3.7 d for Ex-Cell 420 and Grace’s media, respectively. When 60% confluence occurred, cells were infected by viral inoculums. All biochemical compounds were significantly changed relevant to cellular metabolism due to HearNPV infection. In order to improve its stability, two polymer formulations were used, i.e., formulation A (sodium alginate, gelatin, starch, and molasses) and formulation B (cottonseed kernel extract, Bran, glycerol, boric acid, egg white, and sugar). Formulant A provided high photostability by exhibiting 83.2 ± 3% efficacy and 88.66 ± 2.1% original activities remaining after 72 h UV exposure. Percentage original activity remaining of unformulated HearNPV and formulated mixture of B was 38.66 ± 2.6% and 9.33 ± 1.3%, respectively, after 72 h UV-irradiation. The virulence of the HearNPV proliferated from the Ex-Cell medium was similar to the virulence of wild-type HearNPV with LC50 of 7.7×105 OBs/ml. Formulant A, revealed only 20.0 ± 1% reduction in efficacy while the unformulated virus and formulant B faced a reduction of 90.0 ± 3% and 64.0 ± 2% after 72 h of UVA irradiation. Formulant A thus showed a high potential to protect HearNPVs microparticles against UV-inactivation suggesting a new platform for more efficient biological-management of cotton bollworm (specific name Helicoverpa armigera, genus: Helicoverpa, Lepidoptera: Noctuidae) in vivo.

Combination of Imatinib with Cisplatin and Nutlin-3: Functional and Molecular Effects on Bcr-Abl Positive Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2954-2954
Author(s):  
Ioanna Skorta ◽  
Heiko van der Kuip ◽  
Martin Henkes ◽  
Moshe Oren ◽  
Walter E. Aulitzky
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Cellular Response ◽  
Combined Treatment ◽  
Optimal Growth ◽  
Growth Conditions ◽  
P53 Expression ◽  
Complete Eradication ◽  
Abl Kinase ◽  
Highly Effective

Abstract Imatinib is highly effective in inducing remissions in chronic phase CML. However, complete eradication of the malignant clone by Imatinib monotherapy is rare. This prompted us to explore the efficacy of combination of Imatinib with the DNA damaging agent Cisplatin and with Nutlin-3, a compound which induces p53 accumulation by interfering with its binding to Mdm2. We used a Bcr-Abl positive cell line characterized by the loss of Imatinib sensitivity in the presence of optimal growth factor concentrations. Whereas combined treatment of Imatinib with either Cisplatin or Nutlin-3 in low and nontoxic doses induced ∼15–20% cell death, simultaneous treatment of all three compounds was highly effective (∼50% cell death) even in the presence of optimal growth conditions. Importantly, comparable effects were also seen in CFU assays performed with primary CD34 positive cells from 5 CML patients. To examine the molecular mechanisms of these effects we analyzed p53 dependent cellular response to Cisplatin in our cell line model in the presence or absence of Imatinib and/or Nutlin-3. The active Bcr-Abl kinase caused superinduction of p53 protein after DNA damage. We found both an upregulated p53 accumulation and enhanced p21 and Mdm2 RNA and protein levels upon Cisplatin treatment. This phenomenon could be reversed both by siRNA-mediated inhibition of Bcr-Abl expression and by Imatinib-induced inhibition of Bcr-Abl kinase activity: despite of optimal growth conditions inhibition of Bcr-Abl caused a significant reduction of p53 expression and activation. In the presence of Nutlin-3, p53 expression was significantly upregulated upon Cisplatin treatment both with and without Imatinib. However, Nutlin-3 was not capable to restore the Imatinib-induced blockade of the transcriptional activity of p53 on mdm2. The reduced p53 activation observed in Bcr-Abl positive cells treated with Imatinib was paralleled by a shift from cell cycle arrest to cell death both in the presence and absence of Nutlin selectively in Bcr-Abl positive cells rendering them hypersensitive to Cisplatin. In summary, combined treatment of Imatinib with low concentrations of a DNA damaging agent and a p53 activator is highly effective in vitro. Such combined treatment may prove to be clinically relevant for complete eradication of the malignant clone in CML, provided that conditions are found where it does not affect adversely normal hematopoietic cells in vivo.

Optimal growth temperature for the isolation of Plesiomonas shigelloides, using various selective and differential agars

1991 ◽  
Vol 37 (10) ◽  
pp. 800-802 ◽  
Author(s):  
Anwarul Huq ◽  
Anwari Akhtar ◽  
M. A. R. Chowdhury ◽  
David A. Sack
Keyword(s):  
Growth Temperature ◽  
Optimal Growth ◽  
Growth Conditions ◽  
Aeromonas Species ◽  
Growth Media ◽  
Optimal Growth Temperature ◽  
Environmental Waters ◽  
Temperature Growth ◽  
Plesiomonas Shigelloides ◽  
Incubation Temperatures

The growth characteristics of known strains of Plesiomonas shigelloides were compared with those of Aeromonas species (the major competing species in environmental waters) on plesiomonas differential agar, inositol brilliant green bile salt, and modified salmonella–shigella agar at incubation temperatures of 37, 42, and 44 °C. Using local isolates from clinical and environmental sources, optimal growth conditions, as determined by colony counts and the colony characteristics, plesiomonas differential agar proved to be ideal when incubated at 44 °C. Contrary to earlier recommendations for 48 h incubation, the colonies could be recognized readily after an incubation of 24 h. Key words: Plesiomonas, growth temperature, growth media.

Establishment of a New Cell Line from Maxillary Sinus Carcinoma

1980 ◽  
Vol 89 (1) ◽  
pp. 24-28 ◽  
Author(s):  
Tadashi Nakashima ◽  
Kazumi Makishima ◽  
Ikuichiro Hiroto
Keyword(s):  
Maxillary Sinus ◽  
Cell Line ◽  
Single Cells ◽  
Latency Period ◽  
Soft Agar ◽  
Culture Cell ◽  
Population Doubling ◽  
Tissue Culture Cell ◽  
Population Doubling Time

We succeeded in deriving a long-term tissue culture cell line from human maxillary sinus carcinoma. This cell line, designated as MC, was passaged 100 times in vitro over a period of 18 months. The cells are globular in shape, grow as single cells in the culture medium, and the mean population doubling time is about 12 hours. The plating efficiency rate in soft agar is 78% and chromosomal analysis revealed the modal chromosome number to be between 47 and 51. These MC cells were transplanted into five nude mice, all of which developed a tumor after a latency period of 5 to 8 days and died within 39 days. Complete autopsy of all mice revealed no metastasis. Histopathological findings of the original and the transplanted tumor tissues showed a remarkable similarity.

Growth Rates of a New Cell Line from Channel Catfish Ovary and Channel Catfish Virus Replication at Different Temperatures

1980 ◽  
Vol 37 (5) ◽  
pp. 871-873 ◽  
Author(s):  
P. R. Bowser ◽  
J. A. Plumb
Keyword(s):  
Cell Line ◽  
Ictalurus Punctatus ◽  
Channel Catfish ◽  
Growth Rates ◽  
Frozen Storage ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Fish Cell ◽  
Channel Catfish Virus ◽  
New Cell Line

Growth rates of a new cell line from the channel catfish, Ictalurus punctatus, were determined at temperatures from 10 to 35 °C. Growth was more rapid at 35 °C with the cells having a population doubling time of 12 h, and cell numbers declined at 10 °C. The cells survived frozen storage at −70 °C for up to 6 mo. Replication of channel catfish virus occurred from 10 to 35 °C. Although it was most rapid at 35 °C, maximum replication occurred at 30 °C.Key words: fish viruses, channel catfish virus, herpesvirus, fish cell cultures, fish diseases, cell growth rates

scholarly journals Establishment and Characterization of a Cell Line (S-RMS1) Derived from an Infantile Spindle Cell Rhabdomyosarcoma with SRF-NCOA2 Fusion Transcript

2021 ◽  
Vol 22 (11) ◽  
pp. 5484
Author(s):  
Marta Colletti ◽  
Angela Galardi ◽  
Evelina Miele ◽  
Virginia Di Paolo ◽  
Ida Russo ◽  
...  
Keyword(s):  
Whole Genome Sequencing ◽  
Cell Line ◽  
Genome Sequencing ◽  
Gene Fusion ◽  
Spindle Cell ◽  
Population Doubling ◽  
Genetic Profile ◽  
Whole Genome ◽  
A Cell

Background: Spindle cell rhabdomyosarcoma (S-RMS) is a rare tumor that was previously considered as an uncommon variant of embryonal RMS (ERMS) and recently reclassified as a distinct RMS subtype with NCOA2, NCOA1, and VGLL2 fusion genes. In this study, we established a cell line (S-RMS1) derived from a four-month-old boy with infantile spindle cell RMS harboring SRF-NCOA2 gene fusion. Methods: Morphological and molecular characteristics of S-RMS1 were analyzed and compared with two RMS cell lines, RH30 and RD18. Whole genome sequencing of S-RMS1 and clinical exome sequencing of genomic DNA were performed. Results: S-RMS1 showed cells small in size, with a fibroblast-like morphology and positivity for MyoD-1, myogenin, desmin, and smooth muscle actin. The population doubling time was 3.7 days. Whole genome sequencing demonstrated that S-RMS1 retained the same genetic profile of the tumor at diagnosis. A Western blot analysis showed downregulation of AKT-p and YAP-p while RT-qPCR showed upregulation of endoglin and GATA6 as well as downregulation of TGFßR1 and Mef2C transcripts. Conclusion: This is the first report of the establishment of a cell line from an infantile spindle cell RMS with SRF-NCOA2 gene fusion. S-RMS1 should represent a useful tool for the molecular characterization of this rare and almost unknown tumor.

Establishment and characterization of a fibroblast cell line derived from Texel sheep

2009 ◽  
Vol 87 (3) ◽  
pp. 485-492 ◽  
Author(s):  
Linfeng F. Li ◽  
Weijun J. Guan ◽  
Han Li ◽  
Xueyan Z. Zhou ◽  
Xiujuan J. Bai ◽  
...  
Keyword(s):  
Cell Line ◽  
Fluorescent Proteins ◽  
Transfection Efficiency ◽  
Chromosome Analysis ◽  
Fibroblast Cell ◽  
Population Doubling ◽  
Established Cell Line ◽  
Population Doubling Time ◽  
Fibroblast Cell Line ◽  
Plasmid Transfection

A Texel sheep ear marginal tissue fibroblast cell line (named TSF19) was successfully established by using a primary explant technique and cell cryoconservation technology. TSF19 cells were adherent, with a population doubling time of 24.9 h. Chromosome analysis showed that >90% of cells were diploid prior to cell passage 4. Isoenzyme analyses of lactate dehydrogenase and malate dehydrogenase showed that the TSF19 cells had no cross-contamination with other species. Tests for cell line contamination with bacteria, fungi, or mycoplasmas were also negative. Plasmids encoding the fluorescent proteins pEGFP-N3, pECFP-N1, pDsRed1-N1, and pEYFP-N1 were transfected into cells to study exogenous gene expression in the cells. The plasmid transfection efficiency was between 21.8% and 46.5%. This newly established cell line will not only preserve the genetic resources of the important Texel sheep at the cell level but will also provide a valuable resource for genomic, postgenomic, somatic cloning research.

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