Essential Fatty Acid And Cell Culture: Where We Stand

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!

Phone: +1 418.874.0054Toll Free: 1 877.SILICYCLE (North America only)Fax : +1 418.874.0355

[email protected]

SiliCycle Inc - Worldwide Headquarters2500, Parc-Technologique BlvdQuebec City (Quebec) G1P 4S6CANADA

Culture Omega Portfolio (Purity > 99%)

Name of the acid Product Number Type of PUFA CAS #

Eicosapentaenoic (EPA) 50 mg: EPA-GP-050 100 mg: EPA-GP-100 250 mg: EPA-GP-250

ω 3 10417-94-4

Docosapentaenoic (DPA) 50 mg: DPA-GP-050 100 mg: DPA-GP-100 250 mg: DPA-GP-250

ω 3 24880-45-3

Docosahexaenoic (DHA) 50 mg: DHA-GP-050 100 mg: DHA-GP-100 250 mg: DHA-GP-250

ω 3 6217-54-5

α-Linolenic 50 mg: ALN-CG-050 100 mg: ALN-CG-100 250 mg: ALN-CG-250

ω 3 207-40-1

Linoleic 50 mg: LEI-CG-050 100 mg: LEI-CG-100 250 mg: LEI-CG-250

ω 6 60-33-3

y-Linolenic

Available soon

ω 6 506-26-3

Dihomo-y-linolenic ω 6 1783-84-2

Arachidonic ω 6 506-32-1

Docosatetraenoic ω 6 28874-58-0

Eicosadienoic ω 6 2091-39-6

Docosadienoic ω 6 17735-98-7

Eicosatrienoic ω 3 3435-80-1

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.

Essential Fatty Acid Production And Metabolism

pg = prostaglandinpgi = protacyclin

tx = thromboxaneIt = leukotriene

= less inflammatory

= more inflammatoryα-linolenic acid18:3 ω-3

stearidonic acid18:4 ω-3

eicosatetranoic acid20:4 ω-3

eicosapentaenoic acidEPA 20:5 ω-3

docosapentaenoic acidDPA 22:5 ω-3

docosahexaenoic acidDHA 22:6 ω-3

Omega-3 family

α-linolenic acid18:2 ω-6

γ-linolenic acidGLA 18:3 ω-6

dihomo γ-linolenic acidDGLA 20:3 ω-6

arachidonic acidAA 20:4 ω-6

docosatetraenoic acid22:4 ω-6

docosapentaeonoic acid22:5 ω-6

Omega-6 family

pge1 pgf1atxa1

block It4

pgd2 pge2 pgf2apgi2txa2 Ita4 Itb4

Itc4 Itd4 Ite4

pgd3 pge3 pgf3apgi3 txa3

Ita5 Itb5 Itc5 Itd5

A/J-Ring Neuroprostane17S Resolvins

blocks prostanoids

Sprecher’sShunt

6 desaturase

5 desaturase

4 desaturase

elongase

Essential fatty acid production and metabolism Essential Fatty Acid And Cell Culture: Where Do We StandMammalian cells are routinely cultured using the usual basal medium that is a bicarbonate-buffered isotonic aqueous solution, with a high level of glucose supplemented with vitamins as well as essential amino acids. It is boosted with up to 10% of bovine calf serum and customized with a set of purified growth factors and antibiotics. It was recently highlighted that there is a severe lack in the composition of these cell culture medium [1]. Indeed, they are deficient in one of the paramount cell components: essential fatty acids. These polyunsaturated fatty acids (PUFA) are not just an energy source, they constitute an important part in the membrane composition of every single cell and they are necessary for cell signaling.

At the 2013 meeting of the International Federation for Adipose Therapeutics and Science, a researcher from Université Laval presented a physiological model of adipose tissue and demonstrated the importance of incorporating essential fatty acids in the culture medium. Her conclusions were that:

1. There were only trace-levels of essential fatty acids in all the fetal bovine serums tested;

2. The lipid composition of the cell membranes was not physiological and contained mainly saturated fatty acids;

3. Supplementation with essential fatty acids improved the differentiation of adipocytes;

4. Supplementation with essential fatty acids restored the lipid profile of the cell membrane and vacuoles [1].

Since the work of Georges O. Burr and Mildred M. Burr in 1929 [2], it is known that fat is an essential component of the diet. Nowadays, the importance of a balanced omega 3/6 diet is widely recognized for many health functionalities. Unbalance in essential fatty acids can influence chronic inflammation, cardiovascular diseases, brain development and hormonal signaling. Therefore, why are cells still cultured in environments deprived of essential fatty acids?

Several studies have already shown the importance of essential fatty acids in cell culture. Supplementation with omega-3 fatty acids has been shown to reduce inflammation and increase microglial phagocytosis in a model of multiple sclerosis [3]; to supress the pathological phenotype of human

bronchial epithelial cells in a model of cystic fibrosis [4]; and to protect photoreceptors from oxidation-induced apoptosis in rat retinal neuron cells [5]. These in vitro studies demonstrate that various cell types do benefit from supplementation with essential fatty acids, not only adipocytes.

The beneficial effects of essential fatty acid upplementation have been well documented in vivo as well. For example, clinical studies have demonstrated that a diet rich in omega 3 fatty acids can reduce inflammation in obese pregnant women [6] and reduce the progression of Alzheimer’s disease [7].

There is a wide range of opportunities waiting to be seized! The evidences on the benefits of using Culture Omega™ products are piling up. Many cell types are yet to have their first publication stipulating the benefits of essential fatty acid supplementation.

SiliCycle inc. has developed a fatty acids extraction platform using their proprietary silica column. This process allows to isolate large quantities of PUFA as acids (not esters) with a very high level of purity (99%+). These products, tailored for cell culture, are certified to be exempt of bacteria, mycoplasmas and endotoxins.

Contact SiliCycle Inc. today to learn more about the Culture Omega™ product line and to receive your sample:

Tel: +1 418 874-0054Email: [email protected]

References[1] Ouellette ME, et al. (2013) Comparative study of tissue-engineered adipose substitutes and human native fat through transcriptomics and lipid profiling IFAT meeting

[2] Burr GO, et al. (1929) New deficiency disease produced by the rigid exclusion of fat from the diet J Biol Chem

[3] Chen S, et al. (2014) n-3 PUFA supplementation benefits microglial responses to myelin pathology Sci Rep

[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

[5] Simon MV, et al. (2016) Synthesis of docosahexaenoic acid from eicosapentaenoic acid in retina neurons protects photoreceptors from oxidative stress J Neurochem

[6] Haghiac M, et al. (2015) Dietary Omega-3 Fatty Acid Supplementation Reduces Inflammation in Obese Pregnant Women: A Randomized Double-Blind Controlled Clinical Trial PLoS One

[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