The aim of this study was to investigate the effect of fetal bovine serum FBSsupplemented in culture medium for fibroblast cells derived from porcine ovariantissue.. Problem statement Is
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In 1958, fetal bovine serum (FBS) was successfully tested on fibroblast-like cells as a substitute for embryo extract in growth media, leading to its widespread adoption as a crucial component in cell and tissue culture FBS plays a significant role in various techniques related to cell isolation and the establishment of cell lines, particularly for species such as goats and sheep, as documented in multiple studies (Phelan and May, 2015; Li et al., 2009; Bai et al., 2012; Mehrabani et al., 2016; Liu et al., 2011; Wang et al., 2012).
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2.3.4 Ethical stand on the use of FBS in cell culture medium and recent takes on animal-based sera in cell culturing
The use of fetal bovine serum (FBS) has become increasingly discouraged due to significant scientific and ethical concerns Variability in serum composition from different batches undermines the reproducibility of laboratory experiments worldwide, as highlighted by studies indicating that results can differ even among products from the same producer (Honn et al., 1975; Jochems et al., 2002; van der Valk et al., 2010; Franke et al., 2014) Research has shown that bovine fetuses are sentient and experience pain during blood collection, prompting recommendations for more humane harvesting practices, such as ensuring the fetus is unconscious or using a captive bolt for stunning (Jochems et al., 2002; van der Valk et al., 2004) However, many FBS harvesters continue to disregard these guidelines due to the lack of transparency and weak regulations within the FBS market (Gstraunthaler et al., 2014; Hodgson, 1991; Hodgson, 1993; Hodgson, 1995).
The attempt to develop a serum-free medium has been going on for a long time.
In the 1960s, researchers discovered that supplementing insulin alongside low concentrations of serum could promote cell growth in baby hamster kidney cells, as noted in studies by Lieberman and Ove (1959), Higuchi (1963), and Clarke et al (1970) and Clarke and Stoker.
Since the discovery of nerve growth factor in 1952, numerous growth factors have been identified, including epidermal growth factor, insulin-like growth factor, fibroblast growth factor, platelet-derived growth factor, and transforming growth factor These growth factors, when added to culture mediums, have been shown to enhance cell proliferation However, their effectiveness in promoting cell growth remains significantly lower compared to the impact of serum.
In 1976, three studies identified key components for serum substitutes, including selenite, a combination of transferrin and albumin, and various hormones and growth factors This led to the development of serum-free media tailored for different cell types, utilizing ITS (insulin, transferrin, and selenite) alongside appropriate basal media By 2010, van der Valk and colleagues proposed optimizing chemically defined media as a potential replacement for fetal bovine serum (FBS).
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Content 2: Effect of FBS on fibroblast cells proliferative activity
In this study, fibroblast cells were passaged after 8 days of culturing using the optimal FBS concentration identified in previous research The cells were then subcultured into 12-well plates with three different treatments: 10%, 15%, and 20% FBS.
Cells were cultured in 16 wells at 39 °C with 5% CO2 for 7 days, and harvested on Days 1, 3, 5, and 7 post-subculture The cell counts obtained during these intervals were utilized to calculate the Population Doubling Time (PDT) using the time of cell culture in the exponential phase.
NI: number of cells at the start of the exponential phase
N2: number of cells at the end of the exponential phase
This experiment was replicated 4 times.
The data underwent ANOVA analysis with subsequent Tukey testing, utilizing Minitab 16 software, where a significance level of P