2015
DOI: 10.1002/jat.3111
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Effects of homocysteine on mesenchymal cell proliferation and differentiation during chondrogenesis on limb development

Abstract: High levels of homocysteine (Hcy) are related to an increased risk of the occurrence of congenital anomalies, including limb defects. However, few evaluations about how toxic levels of Hcy affect limb development have been reported. We investigated whether Hcy can affect the cell cycle proteins and proteins involved in mesenchymal cell differentiation during limb development, in a chicken embryo model. Embryos were treated with 20 µmol d-l Hcy/50 µl saline at embryonic day 2 and analyzed at embryonic day 6. Un… Show more

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Cited by 8 publications
(1 citation statement)
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“…Then, the cells were dissociated using 0.25% trypsin for 15 min at 37°C and added to 5% FBS under agitation. Subsequently, the samples were centrifuged at 640 ×g for 10 min and the supernatant was collected [ 30 ], incubated with primary antibodies rabbit anti-cyclin E IgG (1 : 1000, Santa Cruz Biotechnology, USA), mouse anti-p21 IgG (1 : 1000, Santa Cruz Biotechnology, USA), and mouse anti- β III tubulin IgG (1 : 1000, Promega, USA) for 1 h, and then incubated for 45 min with the secondary antibodies Alexa Fluor 568 anti-rabbit IgG and Alexa Fluor 488 anti-mouse IgG (1 : 1000, Life Technologies, USA). Analyses were separately conducted for each antibody in each treatment.…”
Section: Methodsmentioning
confidence: 99%
“…Then, the cells were dissociated using 0.25% trypsin for 15 min at 37°C and added to 5% FBS under agitation. Subsequently, the samples were centrifuged at 640 ×g for 10 min and the supernatant was collected [ 30 ], incubated with primary antibodies rabbit anti-cyclin E IgG (1 : 1000, Santa Cruz Biotechnology, USA), mouse anti-p21 IgG (1 : 1000, Santa Cruz Biotechnology, USA), and mouse anti- β III tubulin IgG (1 : 1000, Promega, USA) for 1 h, and then incubated for 45 min with the secondary antibodies Alexa Fluor 568 anti-rabbit IgG and Alexa Fluor 488 anti-mouse IgG (1 : 1000, Life Technologies, USA). Analyses were separately conducted for each antibody in each treatment.…”
Section: Methodsmentioning
confidence: 99%