LABORATORY EXPERIMENTS-PART 2 BIOCHEMISTRY, PLANT BIOTECHNOLOGY, ANIMAL CELL CULTURE, GENETICS

The main purpose of this book is focused on laboratory practical skills are related to the life sciences including Biochemistry, Microbiology and Biotechnology. In the advanced Biological (life sciences) science practical skills and instrumentations are important to the students who studying life science in the area of Biochemistry microbiology and Biotechnology.

Applications and analysis of hydrolysates in animal cell culture

Animal cells are used in the manufacturing of complex biotherapeutic products since the 1980s. From its initial uses in biological research to its current importance in the biopharmaceutical industry, many types of culture media were developed: from serum-based media to serum-free to protein-free chemically defined media. The cultivation of animal cells economically has become the ultimate goal in the field of biomanufacturing. Serum serves as a source of amino acids, lipids, proteins and most importantly growth factors and hormones, which are essential for many cell types. However, the use of serum is unfavorable due to its high price tag, increased lot-to-lot variations and potential risk of microbial contamination. Efforts are progressively being made to replace serum with recombinant proteins such as growth factors, cytokines and hormones, as well as supplementation with lipids, vitamins, trace elements and hydrolysates. While hydrolysates are more complex, they provide a diverse source of nutrients to animal cells, with potential beneficial effects beyond the nutritional value. In this review, we discuss the use of hydrolysates in animal cell culture and briefly cover the composition of hydrolysates, mode of action and potential contaminants with some perspectives on its potential role in animal cell culture media formulations in the future.

Marine cyanobacterium Spirulina maxima as an alternate to the animal cell culture medium supplement

Serum is a stable medium supplement for in vitro cell culture. Live cells are used in stem cell research, drug toxicity and safety testing, disease diagnosis and prevention, and development of antibiotics, drugs, and vaccines. However, use of serum in culture involves concerns such as an ethical debate regarding the collection process, lack of standardized ingredients, and high cost. Herein, therefore, we evaluated the possibility of using edible cyanobacterium (Spirulina maxima), which is a nutrient-rich, sustainable, and ethically acceptable source, as a novel substitute for fetal bovine serum (FBS). H460 cells were cultured to the 10th generation by adding a mixture of spirulina animal cell culture solution (SACCS) and FBS to the culture medium. Cell morphology and viability, cell cycle, apoptosis, proteomes, and transcriptomes were assessed. We observed that SACCS had better growth-promoting capabilities than FBS. Cell proliferation was promoted even when FBS was replaced by 50–70% SACCS; there was no significant difference in cell shape or viability. There were only slight differences in the cell cycle, apoptosis, proteomes, and transcriptomes of the cells grown in presence of SACCS. Therefore, SACCS has the potential to be an effective, low-cost, and eco-friendly alternative to FBS in in vitro culture.

Scale-Up Economics for Cultured Meat: Techno-Economic Analysis and Due Diligence

“Cultured meat” technologies aim to replace conventional meat with analogous or alternative bioproducts from animal cell culture. Developers of these technologies claim their products, also known as “cell-based” or “cultivated” meat, will be safer and more environmentally friendly than conventional meat while offering improved farm-animal welfare. To these ends, Open Philanthropy commissioned this assessment of cultured meat’s potential to measurably displace the consumption of conventional meat.Recognizing that the scalability of any cultured-meat products must in turn depend on the scale and process intensity of animal cell production, this study draws on techno-economic analysis and due-diligence perspectives in industrial fermentation and upstream biopharmaceuticals to assess the extent to which animal cell culture could be scaled like a fermentation process.The analysis identifies a number of significant barriers to the scale-up of animal cell culture. Bioreactor design principles indicate a variety of issues associated with bulk cell growth in culture: Low growth rate, metabolic inefficiency, catabolite and CO2 inhibition, and bubble-induced cell damage will all limit practical bioreactor volume and attainable cell density. With existing bioreactor designs and animal cell lines, a significant engineering effort would be required to address even one of these issues.Economic challenges are further examined. Equipment and facilities with adequate microbial contamination safeguards are expected to have high capital costs. Suitable formulations of amino acids and protein growth factors are not currently produced at scales consistent with food production, and their projected costs at scale are likewise high. The replacement of amino-acid media with plant protein hydrolysates is discussed and requires further study.Capital- and operating-cost analyses of conceptual cell-mass production facilities indicate production economics that would likely preclude the affordability of their products as food. The analysis concludes that metabolic efficiency enhancements and the development of low-cost media from plant hydrolysates are both necessary but insufficient conditions for the measurable displacement of conventional meat by cultured meat.

Elucidating the Biological Activity of Fish-Derived Collagen and Gelatine Hydrolysates using Animal Cell Culture-A review

: A large percentage of a fish's weight is generally discarded during fish processing. Reducing waste in products of marine origin is a subject of great interest within the scientific community. Pelagic by-products such as the structural protein collagen, which can be generated during the processing of fish, has been proposed as an alternative to terrestrial, mammalian sources due to advantages including high availability and low risk of zoonotic disease transmission. Gelatine has multiple possible applications, ranging from nutraceutical applications to cosmetics and has the advantage of being generally regarded as safe. In this multidisciplinary review, the chemistry of gelatine and its parent protein collagen, the chemical reactions to generate their hydrolysates and studies on their biological activities using animal cell culture are discussed.