Viability and cell cycle analysis of equine fibroblasts cultured in vitro

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorLima-Neto, J. F.
dc.creatorFernandes, C. B.
dc.creatorAlvarenga, Marco Antonio
dc.creatorGolim, M. A.
dc.creatorLandim-Alvarenga, Fernanda da Cruz
dc.date2013-09-30T18:28:36Z
dc.date2014-05-20T13:42:03Z
dc.date2016-10-25T16:56:58Z
dc.date2013-09-30T18:28:36Z
dc.date2014-05-20T13:42:03Z
dc.date2016-10-25T16:56:58Z
dc.date2010-08-01
dc.date.accessioned2017-04-05T20:43:19Z
dc.date.available2017-04-05T20:43:19Z
dc.identifierCell and Tissue Banking. Dordrecht: Springer, v. 11, n. 3, p. 261-268, 2010.
dc.identifier1389-9333
dc.identifierhttp://hdl.handle.net/11449/14606
dc.identifierhttp://acervodigital.unesp.br/handle/11449/14606
dc.identifier10.1007/s10561-009-9131-6
dc.identifierWOS:000279295000006
dc.identifierhttp://dx.doi.org/10.1007/s10561-009-9131-6
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/861720
dc.descriptionThis experiment aimed to study equine fibroblasts in culture analyzing and the cell cycle and viability of cells pre- and post-freezing. Skin fragments were obtained from 6 horses and cultured in DMEM high glucose + 10% FCS in 5% CO(2) until the beginning of confluence. Two passages were performed before freezing. Cells subjected to serum starvation (0.5% FCS) were analyzed for viability and cell cycle at 24, 48, 72, 96, 120, 144 and 168 h of culture. For the confluent groups, cells were analyzed at the moment they achieved confluence. Cellular viability was assisted with Hoescht 33342 and propidium iodide. The analysis of apoptosis/necrosis and cell cycle was performed using a flow cytometer (FACS Calibur BD(A (R))) after staining the cells with annexin V and propidium iodide. Both optical microscopy and flow cytometry confirmed that cellular viability was similar for serum starvation and confluent groups (average 84%). Similarly, both methods were efficient to synchronize the cell cycle before freezing. However, after thawing, serum starvation, for more than 24 h, was superior to culture for synchronizing cells in G0/G1 (69% x 90%). The results of this experiment indicate that equine fibroblasts can be efficiently cultured after thawing.
dc.languageeng
dc.publisherSpringer
dc.relationCell and Tissue Banking
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectCell culture Fibroblasts
dc.subjectFlow cytometer
dc.subjectCell cycle
dc.subjectAnnexin V
dc.subjectApoptosis
dc.subjectEquine
dc.titleViability and cell cycle analysis of equine fibroblasts cultured in vitro
dc.typeOtro


Este ítem pertenece a la siguiente institución