scholarly journals Evaluation of Cell Doubling Time in C6 and Y79 Cell Lines Based on Seeding Density

2019 ◽  
Vol 2 (4) ◽  
pp. 146-149
Author(s):  
TS Anitha ◽  
Shreyas S Kuduvalli ◽  
O Ramalakshmi ◽  
S Daisy Precilla
Keyword(s):  
Cell Lines ◽  
Doubling Time ◽  
Seeding Density ◽  
Cell Doubling Time

scholarly journals Quantitative analysis of tumour spheroid structure

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Alexander P Browning ◽  
Jesse A Sharp ◽  
Ryan J Murphy ◽  
Gency Gunasingh ◽  
Brodie Lawson ◽  
...  
Keyword(s):  
Cell Lines ◽  
Tumour Microenvironment ◽  
Culture Conditions ◽  
Experimental Models ◽  
Seeding Density ◽  
Modelling Framework ◽  
Tumour Spheroid ◽  
Tumour Spheroids ◽  
Wide Range

Tumour spheroids are common in vitro experimental models of avascular tumour growth. Compared with traditional two-dimensional culture, tumour spheroids more closely mimic the avascular tumour microenvironment where spatial differences in nutrient availability strongly influence growth. We show that spheroids initiated using significantly different numbers of cells grow to similar limiting sizes, suggesting that avascular tumours have a limiting structure; in agreement with untested predictions of classical mathematical models of tumour spheroids. We develop a novel mathematical and statistical framework to study the structure of tumour spheroids seeded from cells transduced with fluorescent cell cycle indicators, enabling us to discriminate between arrested and cycling cells and identify an arrested region. Our analysis shows that transient spheroid structure is independent of initial spheroid size, and the limiting structure can be independent of seeding density. Standard experimental protocols compare spheroid size as a function of time; however, our analysis suggests that comparing spheroid structure as a function of overall size produces results that are relatively insensitive to variability in spheroid size. Our experimental observations are made using two melanoma cell lines, but our modelling framework applies across a wide range of spheroid culture conditions and cell lines.

Experimental results on mammalian cells growing in vitro in deuterated medium for neutron-scattering studies

1977 ◽  
Vol 25 (1) ◽  
pp. 87-94
Author(s):  
J. Murphy ◽  
C. Desaive ◽  
W. Giaretti ◽  
F. Kendall ◽  
C. Nicolini
Keyword(s):  
Mammalian Cells ◽  
Doubling Time ◽  
Deuterium Oxide ◽  
Initial Period ◽  
Lag Phase ◽  
Suspended Cell ◽  
Human Diploid Fibroblasts ◽  
Cell Doubling Time ◽  
Sv 40 Virus

SV-40 virus-transfromed human diploid fibroblasts (2RA) were grown in a monolayer on plastic Petri dishes in an aqueous medium deuterated to different concentrations of deuterium oxide: 10, 20, 30, up to 60%. The cells must be acclimatized to concentrations higher than 20% D2O by stepping them from a lower initial concentration during their exponential growth. The increase of cell doubling time with increasing deuterium concentrations seems to correlate, at least at 20% D20, with an initial period of suspended cell growth (lag-phase), and is qualitatively similar to that previously reported for Escherichia coli.

G alpha(i-2) mediates renal LLC-PK1 growth by a Raf-independent activation of p42/p44 MAP kinase

1997 ◽  
Vol 272 (2) ◽  
pp. F273-F282 ◽  
Author(s):  
T. B. Kinane ◽  
I. Kang ◽  
A. Chu ◽  
S. H. Chin ◽  
L. Ercolani
Keyword(s):  
Cell Growth ◽  
Protein Kinases ◽  
Doubling Time ◽  
Active Form ◽  
Renal Cells ◽  
Downstream Effectors ◽  
Mitogen Activated Protein ◽  
Cell Doubling Time ◽  
Cell Growth And Differentiation ◽  
Maximal Expression

The protooncogene G alpha(i-2) plays a pivotal role in signaling pathways that control renal cell growth and differentiation. Mitogen-activated protein kinases (MAPKs) are potential downstream effectors for G alpha(i-2) in these pathways. In predifferentiated LLC-PK1 renal cells, the temporal maximal expression of G alpha(i-2) coincided with maximal activation of MAPK(p42/p44). By contrast, pertussis toxin treatment of these cells inhibited cell growth and reduced MAPK(p42/p44) activity by 30%. These findings reflected upstream activation of MAPK kinase (MEK1), as transient transfection of cells with a plasmid encoding a constitutively active form of MEK1 increased MAPK(p42/p44) activity and cell growth, whereas treatment with PD-098059, an inhibitor of MEK1 activity, reduced MAPK(p42/p44) activity and cell growth. Expression of a guanosinetriphosphatase (GTPase)-deficient G alpha(i-2) in these cells increased MAPK(p42/p44) activity and correspondingly reduced cell doubling time from 24 to 10 h without altering the activity of Raf-1 or c-Jun/stress-activated protein kinases (SAPKs). By contrast, expression of a GTPase-deficient G alpha(i-3) in these cells reduced both their cell doubling time by 30% and MAPK(p42/p44) activity by 60%. As the known MEKK isoforms (MEKK1, -2, and -3) can also activate SAPKs, these findings suggest the GTP-charged G alpha(i-2) subunit transduces growth signals in renal cells via activation of MAPK(p42/p44) and that such activation may be linked to pathways containing novel MEKK isoforms that preferentially activate MEKs.

104. PROLIFERATION ABILITY, TELOMERASE ACTIVITY AND MOLECULAR CHARACTERIZATION OF PLURIPOTENT CELL LINES FROM IVF AND PARTHENOGENTIC PIG EMBRYOS

2009 ◽  
Vol 21 (9) ◽  
pp. 23
Author(s):  
T. A. L. Brevini ◽  
G. Pennarossa ◽  
L. Attanasio ◽  
B. Gasparrini ◽  
F. Gandolfi
Keyword(s):  
Cell Lines ◽  
Doubling Time ◽  
Inner Cell Mass ◽  
Es Cells ◽  
Cell Mass ◽  
Telomerase Activity ◽  
Self Renewal ◽  
Pluripotency Markers ◽  
Proliferation Ability ◽  
Receptor Beta

Porcine pluripotent ES cell lines are a promising tool for biotechnology, biomedical and developmental biology studies. However, no conclusive results have been obtained to derive genuine ES cells in the pig. Here we compare derivation efficiency of putative ES cells from IVF versus parthenogenetic pig embryos. We describe proliferation ability and doubling time, we study pluripotency markers and telomerase activity (TA) of the cell lines obtained. Pig oocytes were either fertilized in vitro or parthenogenetically activated. Blastocysts were subjected to immuno-surgery. Inner cell mass were plated and outgrowth expansion was monitored daily. Self renewal molecules were studied by RT-PCR and/or immunocytochemistry for up to 42 passages. TA was measured every five passages. The results obtained indicate that stable cell lines can be generated from IVF and parthenogenetic embryos. The latter appeared less resilient to immuno-surgery but demonstrated a higher ability to produce outgrowths. 77% of the parthenogenetic lines vs only 33% of the IVF ones expressed pluripotency markers and displayed high TA. Regardless to their origin, colonies showed a latency growth period in the 48 hours after plating, they grew exponentially between day 3 and 6 and then, proliferation rate was greatly reduced. Doubling time was estimated to be 31.5 hours. In both IVF and parthenogenetic cell lines, positivity for Oct-4, Nanog, Sox-2, Rex-1, SSEA-4, Alkaline phosphatase, TRA-1-81 and STAT3 was detected; no signal for LIF-Receptor beta and gp130 was shown. These results indicate that the main pluripotency network related molecules are expressed in the porcine species, while a classical LIF-Receptor beta- gp130-STAT3 activation pathway does not appear to be involved in the maintenance of self renewal. Finally, every cell lines expressed high TA, which was turned down once cells were induced to differentiate, indicating a physiologically normal control of TA in these cells.

scholarly journals Establishment of two rabbit mammary epithelial cell lines with distinct oncogenic potential and differentiated phenotype after microinjection of transforming genes.

1986 ◽  
Vol 6 (6) ◽  
pp. 1974-1982 ◽  
Author(s):  
I Garcia ◽  
B Sordat ◽  
E Rauccio-Farinon ◽  
M Dunand ◽  
J P Kraehenbuhl ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Doubling Time ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Oncogenic Potential ◽  
Transformed Cell Line ◽  
Epithelial Cell Lines ◽  
Transforming Genes

The goal of this work was to establish an assay for transformation of epithelial cells. Two epithelial cell lines were obtained after microinjecting transforming genes into primary rabbit mammary secretory cells. The cell lines were analyzed for their oncogenic potential and for the maintenance of a differentiated phenotype. A fully transformed cell line, which retained epithelial cell organization, was obtained by coinjecting simian virus 40 DNA and the activated human c-Ha-ras gene. The proliferation rate of these cells was high, with a doubling time of 16 h. Their growth was anchorage independent, and they had lost contact inhibition. The cells were tumorigenic in nude mice, but had no metastatic potential. Both microinjected DNAs were efficiently transcribed and translated, in contrast to the casein genes, which were expressed in primary cells but not in the transformed cell line. An immortalized cell line established after injection with simian virus 40 DNA alone was characterized by a moderate rate of proliferation with a doubling time of approximately 30 h. The growth of these cells was contact inhibited and anchorage dependent. The cells were not tumorigenic in nude mice. The viral DNA was expressed during early passages, as shown by the presence of the large T antigen in cell nuclei, but not at later passages. A high number of lactogenic hormone receptors were found associated with the cell surface. Despite the presence of these receptors, no induction of genes coding for milk proteins was observed after addition of prolactin. These data demonstrate that this assay system can be used to assess the immortalizing and transforming potential of candidate oncogenes in epithelial cells.

scholarly journals The Cultivation of Human Granulosa Cells

2008 ◽  
Vol 51 (3) ◽  
pp. 165-172 ◽  
Author(s):  
Lenka Brůčková ◽  
Tomáš Soukup ◽  
Jiří Moos ◽  
Martina Moosová ◽  
Jana Pavelková ◽  
...  
Keyword(s):  
Growth Factors ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Doubling Time ◽  
Ovarian Follicle ◽  
Vitro Fertilization ◽  
Thecal Cells ◽  
Cell Doubling Time

The major functions of granulosa cells (GCs) include the production of steroids, as well as a myriad of growth factors to interact with the oocyte during its development within the ovarian follicle. Also FSH stimulates GCs to convert androgens (coming from the thecal cells) to estradiol by aromatase. However, after ovulation the GCs produce progesterone that may maintain a potential pregnancy. Experiments with human GCs are mainly focused on the purification of GCs from ovarian follicular fluid followed by FACS analysis or short-term cultivation. The aim of our study was to cultivate GCs for a long period, to characterize their morphology and phenotype. Moreover, we have cultivated GCs under gonadotropin stimulation in order to simulate different pathological mechanisms during folliculogenesis (e.g. ovarian hyperstimulation syndrome). GCs were harvested from women undergoing in vitro fertilization. Complex oocyte-cumulus oophorus was dissociated by hyaluronidase. The best condition for transport of GCs was optimized as short transport in follicular fluid at 37 °C. GCs expansion medium consisted of DMEM/F12, 2 % FCS, ascorbic acid, dexamethasone, L-glutamine, gentamycine, penicillin, streptomycin and growth factors (EGF, bFGF). GCs transported in follicular fluid and cultivated in 2 % FCS containing DMEM/F12 medium supplemented with follicular fluid presented increased adhesion, proliferation, viability and decreased doubling time. Cell viability was 92 % and mean cell doubling time was 52 hrs. We have optimized transport and cultivation protocols for long-term cultivation of GCs.

scholarly journals Hyperglycemia and hyperinsulinemia stimulate cervical cancer cell proliferation in vitro and in vivo study

2019 ◽  
Author(s):  
Miriam Gutiérrez-Gutiérrez ◽  
Ishell Aline Figueroa-Martínez ◽  
Rafael Jurado ◽  
Norma Uribe ◽  
Patricia García-López ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Cell Proliferation ◽  
Cancer Cell ◽  
Doubling Time ◽  
Xenograft Model ◽  
Cervical Cancer Cell ◽  
Diabetic Patients ◽  
Cell Doubling Time

Abstract Background: Diabetes mellitus and malignant tumor are the second and third causes of women death in Mexico. Hyperglycemia, insulin and insulin-like growth factor 1 are the main risk factors involved in cancer development in patient with diabetes. The aim of this study was to evaluate the effect of hyperglycemia and hyperinsulinemia over cell proliferation and tumor growth in cervical cancer. Methods: Cell proliferation, apoptosis and cell cycle of cervical cancer cell lines (HeLa, SiHa and CaSki) in presence of hyperglycemia and/or insulin were evaluated. Xenograft model for cervical cancer was done in diabetic female nu/nu mice; biochemical parameters, body weight, tumoral volume and cell doubling time were evaluated. Results: Hyperglycemia and hyperinsulinemia significantly increase cell proliferation and decreases apoptosis with no change in cell cycle. Insulin treatment increase tumor volume and diminish cell doubling time, this group also developed hyperinsulinemia and in Langerhans pancreatic islet hypertrophy; whereas, hyperglycemic groups show the same effects but in lesser degree than the insulin treated group. Conclusion: Glucose and insulin stimulates both, proliferation and tumoral growth of cervical cancer, so this should be a possible explanation for the low survival of diabetic patients with cervical cancer in compare to non-diabetic patients with cervical cancer.

scholarly journals LDH-A Knockdown: Changes in The LDH Isoenzyme Profile and Variability in Glioma Response

2021 ◽  
Author(s):  
Masahiro Shindo ◽  
Masatomo Maeda ◽  
Ko Myat ◽  
Mayuresh Mane ◽  
Ivan J. Cohen ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Cell Lines ◽  
Nude Mice ◽  
Doubling Time ◽  
Growth Delay ◽  
Ldh Isoenzyme ◽  
Segmentation Analysis ◽  
Major Energy Source ◽  
Lactate Formation

Abstract Background: Lactate metabolism in tumors is now recognized as a major energy source and a major gluconeogenic precursor for many tumors, as well as shown to exhibit signaling properties. There is less information on the role of the LDH/lactate axis in brain tumors, although lactate formation in gliomas is associated with poor survival. Methods: Three murine glioma cell lines (GL261, CT2A, and ALTS1C1) were transduced to knockdown (KD) expression of the murine LDH-A gene. The effects of the LDH-A KD were compared to those in control (NC) cells and tumors. Results: Differences in the expression of LDH-A and LDH-B mRNA, protein, and enzymatic activity were observed in the six cell lines. LDH zymography showed a major difference in LDH subunit distribution between GL261 LDH-A KD and NC tumors, whereas little or no effect of LDH-A KD was observed in CT2A and ALTS1C1 tumors. Tumors LDH-A and LDH-B immunohistochemistry and a Weka segmentation analysis were consistent with isoenzyme patterns and the above analyses. An “inverse” LDH-A/LDH-B staining relationship (high vs low) was observed in many local GL261 tumor regions. In contrast, CT2A tumors showed a more “direct” local LDH-A/LDH-B staining relationship. LDH-A KD prolonged the doubling time of GL261 cells in culture and prevented the formation of subcutaneous flank tumors in immune-competent C57BL/6 mice (GL261 NC tumors had a prolonged growth delay). In nude mice, both LDH-A KD and NC GL261 tumors grew more rapidly than GL261 NC tumors in C57BL/6 mice. No differences between NC and KD cell proliferation (in vitro) and tumor growth in C57BL/6 mice (doubling time) were observed for CT2A and ALTS1C1 cells and tumors, consistent with the absence of a difference in their LDH isoenzyme profiles. Conclusions: These results show the combined impact of a genetic alteration (LDH-A depletion) on the LDH isoenzyme profile, expression of LDH-A vs LDH-B and LDH enzymatic activity, and the immune system (C57BL/6 vs nude mice) on the growth of s.c. located tumors.

Sign in / Sign up

Export Citation Format

Share Document