Lipids

, Volume 46, Issue 8, pp 753–764 | Cite as

Development of a Fish Cell Culture Model to Investigate the Impact of Fish Oil Replacement on Lipid Peroxidation

  • Melissa K. Gregory
  • Hamish W. King
  • Peter A. Bain
  • Robert A. Gibson
  • Douglas R. Tocher
  • Kathryn A. Schuller
Original Article

Abstract

Fish oils are rich in omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA), predominantly 20:5n-3 and 22:6n-3, whereas vegetable oils contain abundant C18-PUFA, predominantly 18:3n-3 or 18:2n-6. We hypothesized that replacement of fish oils with vegetable oils would increase the oxidative stability of fish lipids. Here we have used the long established and easily cultivated FHM cell line derived from the freshwater fish species fathead minnow (Pimephales promelas) to test this hypothesis. The FHM cells were readily able to synthesize 20:5n-3 and 24:6n-3 from 18:3n-3 but 22:6n-3 synthesis was negligible. Also, they were readily able to synthesize 20:3n-6 from 18:2n-6 but 20:4n-6 synthesis was negligible. Mitochondrial β-oxidation was greatest for 18:3n-3 and 20:5n-3 and the rates for 16:0, 18:2n-6, 22:6n-3 and 18:1n-9 were significantly lower. Fatty acid incorporation was predominantly into phospholipids (79–97%) with very little incorporation into neutral lipids. Increasing the fatty acid concentration in the growth medium substantially increased the concentrations of 18:3n-3 and 18:2n-6 in the cell phospholipids but this was not the case for 20:5n-3 or 22:6n-3. When they were subjected to oxidative stress, the FHM cells supplemented with either 20:5n-3 or 22:6n-3 (as compared with 18:3n-3 or saturated fatty acids) exhibited significantly higher levels of thiobarbituric reactive substances (TBARS) indicating higher levels of lipid peroxidation. The results are discussed in relation to the effects of fatty acid unsaturation on the oxidative stability of cellular lipids and the implications for sustainable aquaculture.

Keywords

Aquaculture β-oxidation Cell culture Fish oil replacement Lipid peroxidation Phospholipids Polyunsaturated fatty acids 

Abbreviations

ALA

α-Linolenic acid

ARA

Arachidonic acid

BHT

Butylated hydroxytoluene

CerPCho

Sphingomyelin

DHA

Docosahexaenoic acid

EDTA

Ethylenediamine tetraacetic acid

EPA

Eicosapentaenoic acid

FAF-BSA

Fatty acid free-bovine serum albumin

FAME

Fatty acid methyl esters

FBS

Foetal bovine serum

FHM

Fathead minnow

HP-TLC

High performance-thin layer chromatography

LC-PUFA

Long-chain polyunsaturated fatty acids (carbon chain length ≥C20 with ≥3 double bonds)

LNA

Linoleic acid

NR

Neutral red

OLA

Oleic acid

PAM

Palmitic acid

PBS

Phosphate buffered saline

PBSA

Phosphate buffered saline without Ca2+ or Mg2+

PCR

Polymerase chain reaction

PtdCho

Phosphatidylcholine

PtdEtn

Phosphatidylethanolamine

Ptd2Gro

Cardiolipin

PtdIns

Phosphatidylinositol

PtdOH

Phosphatidic acid

PtdSer

Phosphatidylserine

PUFA

Polyunsaturated fatty acids

SDS

Sodium dodecyl sulphate

STA

Stearic acid

TBA

Thiobarbituric acid

TBARS

Thiobarbituric acid reactive substances

TLC

Thin-layer chromatography

TN

Total neutral lipids

T/V

Trypsin/versene

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Copyright information

© AOCS 2011

Authors and Affiliations

  • Melissa K. Gregory
    • 1
  • Hamish W. King
    • 1
  • Peter A. Bain
    • 1
  • Robert A. Gibson
    • 2
  • Douglas R. Tocher
    • 3
  • Kathryn A. Schuller
    • 1
  1. 1.School of Biological SciencesFlinders UniversityAdelaideAustralia
  2. 2.School of Agriculture, Food and WineUniversity of AdelaideAdelaideAustralia
  3. 3.Institute of AquacultureUniversity of StirlingStirlingUK

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