Culturing of BHK-21 cells in a medium containing adult bovine serum and pituitary extract

Abstract

To achieve a comparable protein and cell production by culturing of BHK-21 cells in monolayer, in a medium containing adult bovine serum (ABS) and in a medium containing fetal bovine serum (FBS), 1.5 times (v/v) more ABS than FBS has to be added. At a volume fraction in the medium of less than 2% ABS, no proliferation is observed. For a fixed protein and cell production, the amount of required protein in a medium containing ABS can be reduced by addition of a protein extract from pituitary glands from calves (PEP). The relative ceil density after 7 days in a culture medium containing 2 or 5% (v/v) ABS is 3 times higher when 0.2 g/liter pituitary extract is added. This indicates a synergistic interaction between components from ABS and from the protein extract. The addition of a pituitary extract to a medium with ABS also induces a prolonged cell proliferation period. A combined addition to a medium with ABS of 0.02 g/liter PEP and 0.15 g/liter heparin yields almost the same cell density as the addition of 0.2 g/liter PEP alone, indicating the presence of fibroblast growth factor (FGF) in the pituitary extract. However, the enhanced cell production by the addition of a pituitary extract to a medium containing ABS cannot be explained only by the presence in this crude extract of a-or bFGE Abbreviations: ABS -adult bovine serum; BE -protein extract from whole bovine brains; FBS -fetal bovine serum; PEP -protein extract from pituitary glands from calves; PEX -purchased bovine pituitary extract; PDT -population doubling time

Interaction of pepsin with aromatic amino acids and their derivatives immobilized to Sepharose

“…It has been shown that the presence of a thio-ether group is important for introducing selectivity for the given group of proteins [38,49]. N-containing aromatic thiophilic carriers were much less selective for antibodies than regular T-gels [28,50]. Porath and Oscarsson [51] suggested that such immobilized ligands are intermediates between thiophilic and hydrophobic carriers.…”

“…Refs. [25][26][27][28][29][30][31][32][33]), but it could be also employed in the purification of other proteins [26,[33][34][35]. The thiophilic adsorption of proteins is based on an interaction with a sulfone group close to a thioether group of the T-gel ligand.…”

Culturing of BHK-21 cells in a medium, containing adult bovine serum after treatment with hydrophilic silicon particles

“…Obviously, the higher lipid concentration in ABS is not the only reason for the difference between cells cultured in media containing ABS or FBS. For instance a lower concentration of growth factors in ABS may also contribute to this difference [32].…”

Effects of gender-specific adult bovine serum on myogenic satellite cell proliferation, differentiation and lipid accumulation

“…Horse serum is widely used for differentiation (Yaffe et al, 2006) and porcine serum to grow mycoplasma and viruses. Adult bovine serum has been used for BHK cell growth (Strouken et al, 1994) and H pylori culture (Shibayama et al, 2006). Thus, the factors present in serum makes the choice of serum a critical issue in cell culture.…”