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In the present manuscript, the methods required to generate purified cultures of mature adipocytes, as well as stromal vascular cells, from the same isolation are detailed. Also, we describe the in vitro conditions for the dedifferentiation of the isolated mature adipocytes. These two types of cells may be used to reevaluate differences between presently available cellular models for lipogenesis/lipolysis and might provide a new cellular physiological system for studies utilizing the proliferative progeny from mature adipocyte dedifferentiation. Alternative possibilities to the dedifferentiation phenomenon are proposed, as this new area of research is novel.Abbreviations: DMEM -Dulbecco's modified Eagle medium; DMEM/F12 -1:1 ratio; Dulbecco's modified Eagle medium + Ham's F12; FBS -fetal bovine serum; HBSS -Hank's balanced salt solution; HS -horse serum; PBS -phosphate buffered saline, pH 7.08; PSG -pigskin gelatin; SC -satellite cell

Meat animals are unique as experimental models for both lipid metabolism and adipocyte studies because of their direct economic value for animal production. This paper discusses the principles that regulate adipogenesis in major meat animals (beef cattle, dairy cattle, and pigs), the definition of adipose depot-specific regulation of lipid metabolism or adipogenesis, and introduces the potential value of these animals as models for metabolic research including mammary biology and the ontogeny of fatty livers.

Canadian beef consumption is approximately 31 kg per annum, or a third of all meats consumed. Beef is a nutrient-rich food, providing good quality protein, vitamins B-6 and B-12, niacin, iron, and zinc. However, animal fats have gained the reputation of being less healthy. The identification of the anticarcinogenic effects of beef extracts due to the presence of conjugated linoleic acid (CLA) has heightened interest in increasing the amount of CLA deposited in beef. Beef cattle produce CLA and deposit these compounds in the meat; thus, beef consumers can receive bioformed CLA. Beef contains both of the bioactive CLA isomers, namely, cis-9, trans-11 and trans-10, cis-12. The relative content of these CLA isomers in beef depends on the feeds consumed by the animals during production. Feeding cattle linoleic acid-rich oils for extended periods of time increases the CLA content of beef. Depending on the type and relative maturity of the pasture, beef from pasture-fed cattle may have a higher CLA content than beef from grain- or silage-fed cattle. In feedlot animals fed high-grain diets, inclusion of dietary oil along with hay during both the growth and finishing phases led to an increase in CLA content from 2.8 to 14 mg/g beef fat, which would provide 77 mg CLA in an 85-g serving of beef. The CLAs appear to be concentrated in intramuscular and subcutaneous fat of beef cattle, with the CLA trans-10, cis-12 isomer being greater in the subcutaneous fat.

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The deposition of i.m. fat, or marbling, in cattle is recognized as a desirable carcass trait in North American beef grading schemes. In order to investigate the relationship between degree of marbling and fatty acid composition of whole bovine muscle, we extracted the total lipid from pars costalis diaphragmatis (PCD) (n = 23) and longissimus (n = 36) muscles from Wagyu crossbred cattle that were assigned Canadian Grading Agency marbling scores ranging from 1 to 8 on an inverse 10-point scale (i.e., a score of 1 indicated "very abundant" marbling and a score of 10 would be assigned to a carcass "devoid" of marbling). Fatty acid methyl esters (FAME) of the total lipid and triacylglycerol fractions were resolved and quantified through GLC. Marbling scores were negatively associated with total lipid from both PCD (r = -.57, P < .01) and longissimus (r = -.80, P < .001). Differences between PCD and longissimus were found for almost all FAME studied from both lipid fractions, but no differences (P > .05) were seen when the monounsaturated:saturated fatty acid (MUFA/SFA) ratios were compared. Heifers had higher (P < .05) oleic acid content and lower (P < .05) palmitic acid content in lipid extracted from both muscles, resulting in higher (P < .05) MUFA/SFA ratios than those for steers. The relative amount of myristic acid increased as the lipid content (total lipid and triacylglycerol) increased in either longissimus (r values from .48 to .55; n = 36; P < .01) or PCD muscles (r from .67 to .76; n = 23; P < .001). The relative amount of linoleic acid (cis-9, cis-12 isomer) from total lipid was negatively associated with all chemical measurements of lipid from the longissimus (r from -.52 to -.64; n = 36; P < .001) and PCD muscles (r from -.75 to -.85; n = 23; P < .001). This association was not significant (P > .1) for either muscle when linoleic acid from the triacylglycerol fraction was examined, suggesting the negative association between this fatty acid and lipid content was due to a dilution of membrane phospholipids with increasing triacylglycerol. Indices of fatty acid elongase activity, calculated from FAME data, implicated the balance between this enzyme activity and fatty acid synthase as a source of variation between animals displaying various degrees of marbling and worthy of further investigation to better understand the process of marbling fat deposition in beef cattle.

The value of sunflower seed (SS) in finishing diets was assessed in two feeding trials. In Exp. 1, 60 yearling steers (479 +/- 45 kg) were fed five diets (n = 12). A basal diet (DM basis) of 84.5% steam-rolled barley, 9% barley silage, and 6.5% supplement was fed as is (control), with all the silage replaced (DM basis) with rolled SS, or with grain:silage mix replaced with 9% whole SS, 14% whole SS, or 14% rolled SS. Liver, diaphragm, and brisket samples were obtained from each carcass. In Exp. 2, 120 yearling steers (354 +/- 25 kg) were fed corn- or barley-based diets containing no SS, high-linoleic acid SS, or high-oleic acid SS (a 2 x 3 factorial arrangement, n = 20). Whole SS was included at 10.8% in the corn-based and 14% in the barley-based diets (DM basis). In Exp. 1, feeding whole SS linearly increased DMI (P = 0.02), ADG (P = 0.01), and G:F (P = 0.01). Regression of ME against level of whole SS indicated that SS contained 4.4 to 5.9 Mcal ME/kg. Substituting whole for rolled SS did not significantly alter DMI, ADG, or G:F (8.55 vs. 8.30 kg/d; 1.36 vs. 1.31 kg; and 0.157 vs. 0.158, respectively). Replacing the silage with rolled SS had no effect on DMI (P = 0.91) but marginally enhanced ADG (P = 0.10) and improved G:F (P = 0.01). Dressing percent increased linearly (P = 0.08) with level of SS in the diet. Feeding SS decreased (P < 0.05) levels of 16:0 and 18:3 in both diaphragm and subcutaneous fats, and increased (P = 0.05) the prevalence of 18:1, 18:2, cis-9,trans-11-CLA and trans-10,cis-12-CLA in subcutaneous fat. In Exp. 2, barley diets supplemented with high-linoleic SS decreased DMI (P = 0.02) and ADG (P = 0.007) by steers throughout the trial, whereas no decrease was noted with corn (interaction P = 0.06 for DMI and P = 0.01 for ADG). With barley, high-linoleic SS decreased final live weight (554 vs. 592 kg; P = 0.01), carcass weight (329 vs. 346 kg; P = 0.06), and dressing percent (58.5 vs. 59.4%; P = 0.04). Steers fed high-linoleic SS plus barley had less (P < 0.05) backfat than those fed other SS diets. No adverse effects of SS on liver abscess incidence or meat quality were detected. Although they provide protein and fiber useful in formulating finishing diets for cattle, and did improve performance in Exp. 1, no benefit from substituting SS for grain and roughage was detected in Exp. 2. Because of unexplained inconsistencies between the two experiments, additional research is warranted to confirm the feeding value of SS in diets for feedlot cattle.

The effect of breed and diet on insulin response to glucose challenge and its relation to intramuscular fat deposition was determined in 36 steers with 12 each of greater than 87% Wagyu (referred to as Wagyu), Wagyu x Limousin, and Limousin breeds. Weaned steers were blocked by weight into heavy, medium, and light calves and placed in six pens with two pens per weight type and with two steers of each breed per pen. Three pens with steers from each weightclass were fed backgrounding and finishing diets for 259 d, while the other three pens were fed the same diets where 6% of the barley grain was replaced with sunflower oil. Prior to initiation of the finishing phase of the study the intravenous glucose tolerance test (VGTIT) was conducted in all steers. Once steers were judged as carrying adequate 12th-rib fat, based on weight and days on feed, they were harvested and graded and samples of the longissimus muscle were procured for determination of fat content and fatty acid composition. Dietary oil improved (P = 0.011; 0.06) ADG and feed conversion efficiency of steers during the latter part of backgrounding and only ADG during early part ofthe finishing period. Generally percent kidney, pelvic, and heart fat was the only adiposity assessment increased (P = 0.003) by dietary oil. The IVGTT results indicated that insulin response to intravenous glucose was lower in Limousin steers than in Wagyu steers. Dietary oil decreased (P = 0.052) fasting plasma insulin concentration in Wagyu steers compared with Limousin steers. The correlation coefficients among the IVGTT measures and intramuscular fat content or marbling score were less than 0.4, and only a negative trend existed between fasting insulin and USDA marbling scores. However, the carcasses of the Wagyu steers graded US Choice, and 66% of the Wagyu carcasses graded US Prime, which were substantially better than the quality grades obtained for the carcasses from the other breed types. Dietary oil did not affect muscle fat content but increased (P = 0.01) conjugated linoleic acid (CLA) concentrations by 339%. Results indicated that IVGTT measures were not appropriate indices of marbling potential in cattle and that dietary oil can enhance CLA content of beef.