Growth factors from neural tissues have been described as potent mitogens for a wide variety of mesoderm- and ectoderm-derived cells in vitro. We used porcine brain extract for in vitro testing of proliferation properties on primary ovarian cells, uterine cells, and cardiomyocytes in culture as well as for BHK-21 [C-13] cell line. The addition of this extract accelerates proliferation in all examined cultures. It also lowers serum requirement and shortens the cultivation period for BHK-21 [C-13] cells. Fibroblast growth factors from brain of different species, but not porcine, are already characterized and their proliferative effect proved. Therefore, we purified, determined, and confirmed the presence of basic fibroblast growth factor in porcine brain extract by Western blot analysis and showed its biological activity on BHK-21 [C-13] cells. 相似文献
In this study the effects of ammonium and lactate on a culture of channel catfish ovary (CCO) cells were examined. We also
made investigation on the influence of glutamine, since our previous research revealed that this amino acid stimulated CCO
cell growth more than glucose in a concentration-dependent manner. The effect of ammonium in cell culture included the considerable
decrease in cell growth rate with eventual growth arrest as well as the retardation of glucose consumption. At ammonium concentrations
above 2.5 mM, the cells displayed specific morphological changes. The effect of lactate was different to that of ammonium
since the cell growth rate was progressively decreasing with the increase of lactate concentration, whereas the glucose consumption
rate remained almost unchanged. Besides that, it was found that lactate was steadily eliminated from the culture medium when
its initial concentration was relatively high. The influence of glutamine on CCO cell propagation showed that nutrient requirements
of this cell line were mainly dependent on glutamine rather than glucose. The increase in glutamine concentration led to the
increase in cell growth rate and consequent ammonia accumulation while the glucose utilization and lactate production were
reduced. Without glutamine in culture medium cell growth was arrested. However, the lack of glucose reversed the stimulating
effect of glutamine by decreasing cell growth rate and affecting amino acid utilization. 相似文献
In this work we describe the adaptation of channel catfish ovary (CCO) cell line to commercially available Ultra Culture serum-free medium by gradual reduction of serum concentration from 10 to 0 %. With this approach we obtained CCO cells fully adapted to serum-free conditions in 32 days. Growth, nutritional and morphological characteristics of these cells remained unchanged when compared to the control group kept in the presence of serum. Additionally, three commercially available protein hydrolysates were tested for the effects on growth performance of the newly serum-free adapted CCO cells. Supplementation with wheat gluten hydrolysate resulted in growth similar to serum free medium solely, while yeast and soy hydrolysates showed inhibitory effects on the cell growth. Taken together, the successful adaptation of CCO cells to serum-free conditions indicates their potential to be used in cytotoxicity assays when serum omission is demanded or for developing serum free bioprocesses using CCO cells. However, a more extended study on nutrient supplementation is still required to further boost the cell growth in a serum free culture. 相似文献
Food proteins from different sources can provide beneficial effects on human health by releasing the bioactive peptides that are integral part of their native structure. In this study, we tested the biological potential of hempseed protein hydrolysates (HPHs) obtained from hempseed cake protein isolate. The HPHs were prepared by enzyme hydrolysis using three different proteases of microbial origin: Alcalase®, Neutrase® and Protamex®. The antioxidant activity of the obtained hydrolysates was determined by oxygen radical absorbance capacity (ORAC) assay, while the proliferative effects on normal (HaCaT) and cancer (HeLa) cells were determined by the CellTiter 96® AQueous One Solution Reagent (MTS) assay. HPHs showed dose-dependent antiproliferative effects on HeLa cells and stimulatory effects on the proliferation of HaCaT cells. HPH obtained by Neutrase® (HPH-N) showed the highest antioxidant activity expressed as an ORAC value. The protective effect of HPH-N on H2O2-induced oxidative stress in normal and cancer cells was evaluated and 1 mg/mL of HPH-N significantly reduced the formation of intracellular reactive oxygen species (ROS) in both cell lines. The obtained results indicate the benefits of HPHs as potential natural antioxidants for the food industry and contribute to the growing trend of utilizing hempseed by-products.
A novel, disposable-bag bioreactor system that uses wave action for mixing and transferring oxygen was evaluated for BHK 21
C13 cell line growth and Aujeszky’s disease virus (ADV) production. Growth kinetics of BHK 21 C13 cells in the wave bioreactor
during 3-day period were determined. At the end of the 3-day culture period and cell density of 1.82 × 106 cells ml-1, the reactor was inoculated with 9 ml of gE- Bartha K-61 strain ADV suspension (105.9 TCID50) with multiplicity of infection (MOI) of 0.01. After a 144 h incubation period, 400 ml of ADV harvest was obtained with titre
of 107.0 TCID50 ml−1, which corresponds to 40,000 doses of vaccine against AD. In conclusion, the results obtained with the wave bioreactor using
BHK 21 C13 cells showed that this system can be considered as suitable for ADV or BHK 21 C13 cell biomass production. 相似文献
The growth characteristics and influence of glucose and glutamine on the growth and maintenance of channel catfish ovary (CCO)
cells were investigated. Besides glutamine, amino acids threonine, arginine, methionine and serine were found to be essential
for CCO cell growth. In the glucose-free medium, glutamine is utilized as energy source and no cell growth limitation was
observed. However, the lack of glutamine in culture medium did not stimulate CCO cells to efficient glucose consumption. When
both glucose and glutamine were deficient, cell growth was also observed suggesting no rigorous nutritional requirements.
Obtained results are useful for further understanding of culture processes using CCO cells. 相似文献