Effect of serum starvation and chemical inhibitors on cell cycle synchronization of canine dermal fibroblasts

Khammanit, R.; Chantakru, Sirirak; Kitiyanant, Yindee; Saikhun, Jumnian

doi:10.1016/j.theriogenology.2008.02.015

Cited by 69 publications

(78 citation statements)

References 27 publications

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“…The contrasting outcomes in these studies might be the consequence of the treatment methods, species, or cell type. Khammanit et al (2008) reported that serum starvation did not increase the rate of apoptotic cells; by contrast, in the present study, the rate of apoptosis after 48 h serum starvation was higher than found for contact inhibition or roscovitine treatments. We also examined the relative levels of expression of apoptosis-related genes, BAX and BCL-2.…”

Section: Discussioncontrasting

confidence: 99%

Production of hGFAP-DsRed transgenic Guangxi Bama mini-pigs via somatic cell nuclear transfer

Lü

Yang

et al. 2015

Genet. Mol. Res.

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ABSTRACT. The mini-pig is a useful animal model for human biomedical research due to its physiological similarity to humans and the ease of handling. In order to optimize the efficiency of production of transgenic Bama mini-pigs through somatic cell nuclear transfer (SCNT), we examined the effects of contact inhibition, roscovitine treatment, and serum starvation on the cell cycle synchronization and transgenic cloned embryo development in vivo and in vitro after nuclear transfer. The analysis showed that the rates of G0/G1 stage cells in the contact inhibition (92.11%) and roscovitine treatment groups (89.59%) were significantly higher than in serum starvation group (80.82%). A higher rate of apoptosis was seen in the serum starvation group (14.13%) compared to the contact inhibition and roscovitine treatment groups (6.71 and 2.46% respectively, P < 0.05). There was a significant decrease in blastocyst yield in the serum starvation group (14.19%) compared to the roscovitine treatment and contact inhibition groups (21.31 and 20.32% respectively, P < 0.05). A total of 1070 transgenic cloned embryos derived from the three treatment groups were 16286 L.L. Hu et al.©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (4): 16285-16296 (2015) transferred to surrogate sows; one pregnancy was established and three embryos from the roscovitine treatment group successfully completed gestation. These results indicate that the roscovitine treatment was more effective at synchronizing transgenic kidney cells in Bama mini-pigs and allowed reconstructed embryos to develop to full term.

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Section: Discussioncontrasting

confidence: 99%

Production of hGFAP-DsRed transgenic Guangxi Bama mini-pigs via somatic cell nuclear transfer

Lü

Yang

et al. 2015

Genet. Mol. Res.

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“…This reversible effect of colchicine on the cytokinesis of eukaryotic cells has been previously observed using low drug concentrations. Treatment of canine epidermal fibroblasts with 2.5 × 10 −4 mM of colchicine blocked the cell cycle, although removal of the drug and replacement with fresh medium completely restored cell division (Khammanit et al 2008). This reversibility in the effect observed with colchicine treatment on different cell types may be the direct result of the intrinsic dynamic nature of microtubule assembly-disassembly (Desai and Mitchison 1997).…”

Section: Discussionmentioning

confidence: 95%

Colchicine treatment reversibly blocks cytokinesis but not mitosis in Trypanosoma cruzi epimastigotes

Potenza

Téllez-Iñón

2014

Parasitol Res

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This work analyzes the effect of the alkaloid colchicine on the growth of Trypanosoma cruzi epimastigotes, using immunofluorescence microscopy and flow cytometry techniques. We found that colchicine reversibly inhibited cytokinesis but not synthesis or segregation of nuclear and kinetoplastid DNA, in a concentration-dependent manner. We showed that, once colchicine was removed from the growth medium, cytokinesis was restored but abnormal segregation of kinetoplasts and nuclei generated zoids and parasites with two nuclei and one kinetoplast, among other aberrant cells. After drug removal, we also observed a few anucleated cells carrying two kinetoplasts in a stage compatible with the end of cytokinesis. The anomalous subcellular localization of the kinetoplast and flagellum observed in treated parasites suggests that the effect of colchicine and its interaction with T. cruzi microtubules is cell cycle dependent. The crosstalk between nuclear and kinetoplastid mitosis and its incidence on flagellum growth and parasite cell division regulation are discussed.

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“…Densities of 1.56 × 10 4 cells/cm 2 and above represent confluent and post-confluent cultures respectively. The cultures were synchronized with serum starvation for 24 h (Khammanit et al 2008) followed by treatment with medium containing 10% (v/v) FBS with or without TGF-β (1 ng/mL) for an additional 24 h. The conditioned media were collected and centrifuged at 1000 g for 5 min to remove any cellular debris and stored at -20°C until used. All experiments were performed at least three times and carried out in triplicate.…”

Section: Tissue Samples and Cell Culturesmentioning

confidence: 99%

Cell density-dependent stimulation of PAI-1 and hyaluronan synthesis by TGF-β in orbital fibroblasts

Galgoczi

Jeney

Gazdag

et al. 2016

Journal of Endocrinology

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During the course of Graves’ orbitopathy (GO), orbital fibroblasts are exposed to factors that lead to proliferation and extracellular matrix (ECM) overproduction. Increased levels of tissue plasminogen activator inhibitor type 1 (PAI-1 (SERPINE1)) might promote the accumulation of ECM components. PAI-1 expression is regulated by cell density and various cytokines and growth factors including transforming growth factorβ(TGF-β). We examined the effects of increasing cell densities and TGF-β on orbital fibroblasts obtained from GO patients and controls. Responses were evaluated by the measurement of proliferation, PAI-1 expression, and ECM production. There was an inverse correlation between cell density and the per cell production of PAI-1. GO orbital, normal orbital, and dermal fibroblasts behaved similarly in this respect. Proliferation rate also declined with increasing cell densities. Hyaluronan (HA) production was constant throughout the cell densities tested in all cell lines. In both GO and normal orbital fibroblasts, but not in dermal fibroblasts, TGF-β stimulated PAI-1 production in a cell density-dependent manner, reaching up to a five-fold increase above baseline. This has been accompanied by increased HA secretion and pericellular HA levels at high cell densities. Increasing cell density is a negative regulator of proliferation and PAI-1 secretion both in normal and GO orbital fibroblasts; these negative regulatory effects are partially reversed in the presence of TGF-β. Cell density-dependent regulation of PAI-1 expression in the orbit, together with the local cytokine environment, may have a regulatory role in the turnover of the orbital ECM and may contribute to the expansion of orbital soft tissue in GO.

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Effect of serum starvation and chemical inhibitors on cell cycle synchronization of canine dermal fibroblasts

Cited by 69 publications

References 27 publications

Production of hGFAP-DsRed transgenic Guangxi Bama mini-pigs via somatic cell nuclear transfer

Production of hGFAP-DsRed transgenic Guangxi Bama mini-pigs via somatic cell nuclear transfer

Colchicine treatment reversibly blocks cytokinesis but not mitosis in Trypanosoma cruzi epimastigotes

Cell density-dependent stimulation of PAI-1 and hyaluronan synthesis by TGF-β in orbital fibroblasts

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