Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts

Abstract

BackgroundAirway fibroblasts have become a critical addition to all facets of structural lung tissue changes such as in human asthma and chronic obstructive pulmonary disease, but little is known about their role in the equine recurrent airway obstruction, a disease that resembles to the human asthma. Since the equine bronchial fibroblasts (EBF) have not been isolated and characterized yet, the use of defined medium was investigated.ResultsPrimary EBF were cultured on non-collagen coated flasks without serum or in the presence of fetal bovine serum (FBS) or horse serum (HS) or in serum depleted medium. EBF cultured in serum-free basal media and those serum deprived were not able to proliferate and even exhibited considerable cell death. In media containing FBS or HS, proliferation of the cells was reproducible between different primary cultures and cells demonstrated expression of vimentin. Large variations were found in the ability of FBS and HS to support growth and differentiation of EBF in monolayer culture. Indications of growth-promoting actions, increasing passage number as well as maintaining fibroblast morphology were found rather in FBS than in HS. EBF culturing in HS needed longer doubling and confluence time. The protein content of the cell pellets was higher in EBF cultured in medium containing HS than FBS. Alpha-smooth muscle actin seemed to be less expressed in EBF cultured in medium containing FBS than those in HS.ConclusionsIn sum, serum addition to basal EBF medium enhanced EBF differentiation into myofibroblasts, and these findings are useful to develop in vitro fibroblast culture models that mimic in vivo physiological processes and to study airway disease mechanisms and remodeling.

In vitro spontaneous differentiation of human breast cancer stem cells and methods to control this process

“…FBS, which contains a pool of undetermined factors and components, caused the differentiation of BCSCs. Although FBS is an essential component of cell culture medium, it has been shown to cause differentiation of certain kinds of stem cells such as neural and embryonic, as well as germ cells (Bettiol et al, 2007;Franke et al, 2014;Hung and Young, 2006;Tateishi et al, 2008;Zahir et al, 2009). …”

A simplified protocol for the isolation and culture of cardiomyocytes and progenitor cells from neonatal mouse ventricles

“…As we did not characterize these structures for any other markers beside c-KIT, we could not confirm their cardiosphere nature. Fibroblasts that detach during trypsin treatment are present around the spheroids that, influenced by serum (Franke et al, 2014), tend to proliferate and generate confluent cultures of non-myocytes (Fig. 3D3).…”

“…To reduce the non-myocyte population during sub-culture with enzymatic dissociation, we included hydrocortisone as a supplement to the protocol. The cultures response to 1 M hydrocortisone was tested in high serum concentration as HS aids proliferation of fibroblasts (Franke et al, 2014). Hydrocortisone inhibits fibroblast proliferation (Holden and Adams, 1957) while inducing the expression of COX2 (Ptgs2) in cardiomyocyte cultures (Sun et al, 2008) and COX2 activity in cardiomyocytes is cardio protective (Inserte et al, 2009).…”

In vitro evaluation of the cytotoxicity and modulation of mechanisms associated with inflammation induced by perfluorooctanesulfonate and perfluorooctanoic acid in human colon myofibroblasts CCD-18Co

“…DMSO (0.5% v/v) was used to treat the control cells. As FBS is required for growth and proliferation of the fibroblasts (Franke et al, 2014), deprived-FBS medium was discarded for this assay. Every 24 h of incubation (up to 96 h), the culture medium was removed and the cells were washed with PBS.…”