Essential Fatty Acid and Cell Culture: Where We Stand

Essential Fatty Acid And Cell Culture: Where We Stand

Phone: +1 418.874.0054 Toll Free: 1 877.SILICYCLE (North America only) Fax : +1 418.874.0355 [email protected] www.SiliCycle.com

SiliCycle Inc - Worldwide Headquarters 2500, Parc-Technologique Blvd Quebec City (Quebec) G1P 4S6 CANADA

Tissue engineering aims at creating relevant human in vitro models for evaluation of drugs or for transplantation. Those models are intended to be as physiological as possible. However, essential fatty acids are currently ignored in the process. Essential for proper membrane fluidity, these lipidic building-blocks are also implicated in several cellular processes, including cell signaling. In fact, the beneficial effects of a proper omega-3 diet were clearly established in several clinical studies for a large array of pathological conditions including cardiovascular diseases, diabetes, chronic inflammation and neurodegenerative diseases. Ultimately, these in vivo effects are orchestrated at the cellular level; hence supplementation with essentials fatty acids is becoming paramount for cell culture models as it started to emerge. Conditions are met for cell biology to integrate fatty acids in the culture medium and enter the lipidomic era!

© SiliCycle inc. 2017 EssentialEssential Fatty fatty Acid acid Production production And and Metabolism metabolism Essential Fatty Acid And Cell Culture: Where Do We Stand pg = prostaglandin tx = thromboxane pgi = protacyclin It = leukotriene Mammalian cells are routinely cultured using the usual basal Omega-3 family Omega-6 family medium that is a bicarbonate-buffered isotonic aqueous = less inflammatory solution, with a high level of glucose supplemented with α-linolenic acid = more inflammatory α-linolenic acid vitamins as well as essential amino acids. It is boosted with 18:3 ω-3 18:2 ω-6 up to 10% of bovine calf serum and customized with a set 6 desaturase of purified growth factors and antibiotics. It was recently highlighted that there is a severe lack in the composition stearidonic acid γ-linolenic acid of these cell culture medium [1]. Indeed, they are deficient 18:4 ω-3 GLA 18:3 ω-6 bronchial epithelial cells in a model of cystic fibrosis [4]; and in one of the paramount cell components: essential fatty to protect photoreceptors from oxidation-induced apoptosis in elongase acids. These polyunsaturated fatty acids (PUFA) are not just an energy source, they constitute an important part in the rat retinal neuron cells [5]. These in vitro studies demonstrate that various cell types do benefit from supplementation with eicosatetranoic acid pge1 pgf1a dihomo γ-linolenic acid membrane composition of every single cell and they are txa essential fatty acids, not only adipocytes. 20:4 ω-3 1 DGLA 20:3 ω-6 necessary for cell signaling. block It4 At the 2013 meeting of the International Federation for The beneficial effects of essential fatty acid upplementation 5 desaturase Adipose Therapeutics and Science, a researcher from have been well documented in vivo as well. For example, clinical studies have demonstrated that a diet rich in omega pgd pge pgf pgd pge pgf Université Laval presented a physiological model of adipose 3 3 3a 2 2 2a 3 fatty acids can reduce inflammation in obese pregnant eicosapentaenoic acid pgi3 txa3 pgi2txa2 Ita4 Itb4 arachidonic acid tissue and demonstrated the importance of incorporating women [6] and reduce the progression of Alzheimer’s EPA 20:5 ω-3 Ita5 Itb5 Itc5 Itd5 Itc4 Itd4 Ite4 AA 20:4 ω-6 essential fatty acids in the culture medium. Her conclusions were that: disease [7]. Sprecher’s elongase Shunt There is a wide range of opportunities waiting to be seized! 1. There were only trace-levels of essential fatty acids in all docosapentaenoic acid docosatetraenoic acid The evidences on the benefits of using Culture Omega™ the fetal bovine serums tested; DPA 22:5 ω-3 22:4 ω-6 products are piling up. Many cell types are yet to have their 4 desaturase 2. The lipid composition of the cell membranes was not first publication stipulating the benefits of essential fatty acid physiological and contained mainly saturated fatty acids; supplementation. docosahexaenoic acid A/J-Ring Neuroprostane docosapentaeonoic acid DHA 22:6 ω-3 17S Resolvins 22:5 ω-6 3. Supplementation with essential fatty acids improved the SiliCycle inc. has developed a fatty acids extraction platform blocks prostanoids differentiation of adipocytes; using their proprietary silica column. This process allows to isolate large quantities of PUFA as acids (not esters) 4. Supplementation with essential fatty acids restored the with a very high level of purity (99%+). These products, of the cell membrane and vacuoles [1]. tailored for cell culture, are certified to be exempt of bacteria, Culture Omega Portfolio (Purity > 99%) lipid profile mycoplasmas and endotoxins. Name of the acid Product Number Type of PUFA CAS # Since the work of Georges O. Burr and Mildred M. Burr in 50 mg: EPA-GP-050 1929 [2], it is known that fat is an essential component of Contact SiliCycle Inc. today to learn more about the Eicosapentaenoic (EPA) 100 mg: EPA-GP-100 ω 3 10417-94-4 250 mg: EPA-GP-250 the diet. Nowadays, the importance of a balanced omega Culture Omega™ product line and to receive your sample: Docosapentaenoic (DPA) 50 mg: DPA-GP-050 3/6 diet is widely recognized for many health functionalities. 100 mg: DPA-GP-100 ω 3 24880-45-3 Tel: +1 418 874-0054 250 mg: DPA-GP-250 Unbalance in essential fatty acids can influence chronic Email: [email protected] Docosahexaenoic (DHA) 50 mg: DHA-GP-050 inflammation, cardiovascular diseases, brain development 100 mg: DHA-GP-100 ω 3 6217-54-5 250 mg: DHA-GP-250 and hormonal signaling. Therefore, why are cells still cultured in environments deprived of essential fatty acids? α-Linolenic 50 mg: ALN-CG-050 100 mg: ALN-CG-100 ω 3 207-40-1 250 mg: ALN-CG-250 Several studies have already shown the importance of Linoleic 50 mg: LEI-CG-050 References 100 mg: LEI-CG-100 ω 6 60-33-3 essential fatty acids in cell culture. Supplementation with 250 mg: LEI-CG-250 omega-3 fatty acids has been shown to reduce inflammation [1] Ouellette ME, et al. (2013) Comparative study of tissue-engineered adipose substitutes and human native fat through transcriptomics and lipid profiling IFAT meeting y-Linolenic ω 6 506-26-3 and increase microglial phagocytosis in a model of multiple sclerosis [3]; to supress the pathological phenotype of human [2] Burr GO, et al. (1929) New deficiency disease produced by the rigid exclusion of fat Dihomo-y-linolenic ω 6 1783-84-2 from the diet J Biol Chem [3] Chen S, et al. (2014) n-3 PUFA supplementation benefits microglial responses to Arachidonic ω 6 506-32-1 myelin pathology Sci Rep

Docosatetraenoic Available soon ω 6 28874-58-0 [4] Njoroge SW, et al. (2012) DHA and EPA reverse cystic fibrosis-related FA abnormalities by suppressing FA desaturase expression and activity J Lipid Res Eicosadienoic ω 6 2091-39-6 [5] Simon MV, et al. (2016) Synthesis of docosahexaenoic acid from eicosapentaenoic acid in retina neurons protects photoreceptors from oxidative stress J Neurochem Docosadienoic ω 6 17735-98-7 [6] Haghiac M, et al. (2015) Dietary Omega-3 Fatty Acid Supplementation Reduces Eicosatrienoic ω 3 3435-80-1 Inflammation in Obese Pregnant Women: A Randomized Double-Blind Controlled Clinical Trial PLoS One Provided in amber ampoules, under vacuum for highest chemical integrity and without stabilizer for unbiased biological testing. Exact weight on the label for precise and convenient dilutions. Tested for biocontaminants. Purity 99%+ (ELSD-MS). Store at -80°C. [7] Wang X, et al. (2015) Effects of n-3 FA supplementation on the release of proresolving lipid mediators by blood mononuclear cells: the OmegAD study J Lipid Res EssentialEssential Fatty fatty Acid acid Production production And and Metabolism metabolism Essential Fatty Acid And Cell Culture: Where Do We Stand pg = prostaglandin tx = thromboxane pgi = protacyclin It = leukotriene Mammalian cells are routinely cultured using the usual basal Omega-3 family Omega-6 family medium that is a bicarbonate-buffered isotonic aqueous = less inflammatory solution, with a high level of glucose supplemented with α-linolenic acid = more inflammatory α-linolenic acid vitamins as well as essential amino acids. It is boosted with 18:3 ω-3 18:2 ω-6 up to 10% of bovine calf serum and customized with a set 6 desaturase of purified growth factors and antibiotics. It was recently highlighted that there is a severe lack in the composition stearidonic acid γ-linolenic acid of these cell culture medium [1]. Indeed, they are deficient 18:4 ω-3 GLA 18:3 ω-6 bronchial epithelial cells in a model of cystic fibrosis [4]; and in one of the paramount cell components: essential fatty to protect photoreceptors from oxidation-induced apoptosis in elongase acids. These polyunsaturated fatty acids (PUFA) are not just an energy source, they constitute an important part in the rat retinal neuron cells [5]. These in vitro studies demonstrate that various cell types do benefit from supplementation with eicosatetranoic acid pge1 pgf1a dihomo γ-linolenic acid membrane composition of every single cell and they are txa essential fatty acids, not only adipocytes. 20:4 ω-3 1 DGLA 20:3 ω-6 necessary for cell signaling. block It4 At the 2013 meeting of the International Federation for The beneficial effects of essential fatty acid upplementation 5 desaturase Adipose Therapeutics and Science, a researcher from have been well documented in vivo as well. For example, clinical studies have demonstrated that a diet rich in omega pgd pge pgf pgd pge pgf Université Laval presented a physiological model of adipose 3 3 3a 2 2 2a 3 fatty acids can reduce inflammation in obese pregnant eicosapentaenoic acid pgi3 txa3 pgi2txa2 Ita4 Itb4 arachidonic acid tissue and demonstrated the importance of incorporating women [6] and reduce the progression of Alzheimer’s EPA 20:5 ω-3 Ita5 Itb5 Itc5 Itd5 Itc4 Itd4 Ite4 AA 20:4 ω-6 essential fatty acids in the culture medium. Her conclusions were that: disease [7]. Sprecher’s elongase Shunt There is a wide range of opportunities waiting to be seized! 1. There were only trace-levels of essential fatty acids in all docosapentaenoic acid docosatetraenoic acid The evidences on the benefits of using Culture Omega™ the fetal bovine serums tested; DPA 22:5 ω-3 22:4 ω-6 products are piling up. Many cell types are yet to have their 4 desaturase 2. The lipid composition of the cell membranes was not first publication stipulating the benefits of essential fatty acid physiological and contained mainly saturated fatty acids; supplementation. docosahexaenoic acid A/J-Ring Neuroprostane docosapentaeonoic acid DHA 22:6 ω-3 17S Resolvins 22:5 ω-6 3. Supplementation with essential fatty acids improved the SiliCycle inc. has developed a fatty acids extraction platform blocks prostanoids differentiation of adipocytes; using their proprietary silica column. This process allows to isolate large quantities of PUFA as acids (not esters) 4. Supplementation with essential fatty acids restored the with a very high level of purity (99%+). These products, of the cell membrane and vacuoles [1]. tailored for cell culture, are certified to be exempt of bacteria, Culture Omega Portfolio (Purity > 99%) lipid profile mycoplasmas and endotoxins. Name of the acid Product Number Type of PUFA CAS # Since the work of Georges O. Burr and Mildred M. Burr in 50 mg: EPA-GP-050 1929 [2], it is known that fat is an essential component of Contact SiliCycle Inc. today to learn more about the Eicosapentaenoic (EPA) 100 mg: EPA-GP-100 ω 3 10417-94-4 250 mg: EPA-GP-250 the diet. Nowadays, the importance of a balanced omega Culture Omega™ product line and to receive your sample: Docosapentaenoic (DPA) 50 mg: DPA-GP-050 3/6 diet is widely recognized for many health functionalities. 100 mg: DPA-GP-100 ω 3 24880-45-3 Tel: +1 418 874-0054 250 mg: DPA-GP-250 Unbalance in essential fatty acids can influence chronic Email: [email protected] Docosahexaenoic (DHA) 50 mg: DHA-GP-050 inflammation, cardiovascular diseases, brain development 100 mg: DHA-GP-100 ω 3 6217-54-5 250 mg: DHA-GP-250 and hormonal signaling. Therefore, why are cells still cultured in environments deprived of essential fatty acids? α-Linolenic 50 mg: ALN-CG-050 100 mg: ALN-CG-100 ω 3 207-40-1 250 mg: ALN-CG-250 Several studies have already shown the importance of Linoleic 50 mg: LEI-CG-050 References 100 mg: LEI-CG-100 ω 6 60-33-3 essential fatty acids in cell culture. Supplementation with 250 mg: LEI-CG-250 omega-3 fatty acids has been shown to reduce inflammation [1] Ouellette ME, et al. (2013) Comparative study of tissue-engineered adipose substitutes and human native fat through transcriptomics and lipid profiling IFAT meeting y-Linolenic ω 6 506-26-3 and increase microglial phagocytosis in a model of multiple sclerosis [3]; to supress the pathological phenotype of human [2] Burr GO, et al. (1929) New deficiency disease produced by the rigid exclusion of fat Dihomo-y-linolenic ω 6 1783-84-2 from the diet J Biol Chem [3] Chen S, et al. (2014) n-3 PUFA supplementation benefits microglial responses to Arachidonic ω 6 506-32-1 myelin pathology Sci Rep

Phone: +1 418.874.0054 Toll Free: 1 877.SILICYCLE (North America only) Fax : +1 418.874.0355 [email protected] www.SiliCycle.com

SiliCycle Inc - Worldwide Headquarters 2500, Parc-Technologique Blvd Quebec City (Quebec) G1P 4S6 CANADA