Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

Background

There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats.

Results

Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection.

Conclusions

Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

Electronic supplementary material

The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.     

Effect of lipid source and bile salts in diet of Atlantic salmon, Salmo salar L., on astaxanthin blood levels

A study was conducted to determine the effects of dietary lipid and bile acids on astaxanthin absorption in Atlantic salmon (Salmo salar L.). Fish with an average weight of 1500 g were fitted with a dorsal aorta cannula and fed diets containing herring oil, soybean lecithin, lard, or herring oil supplemented with taurocholic acid (2.5 g/kg diet). Each fish was fed all of the experimental diets in successive order to minimize the effect of individual variation. At a given time following the feeding of each diet, blood was collected and analyzed for astaxanthin. Soybean lecithin significantly lowered the absorption of astaxanthin compared to fish fed herring oil. A 20% (p < 0.12) increase in blood astaxanthin was observed when the fish were fed the diet supplemented with taurocholic acid. Feeding lard significantly increased the blood astaxanthin level compared to the control group. It appears that altering the micellar structure by stimulating micellar (taurocholic acid) or mixed micellar (lecithin) systems did not increase the apparent absorption of astaxanthin. However, increasing the phospholipid level may have actually decreased the absorption possibly by lowering the astaxanthin solubility in the micelles. The increased apparent absorption of astaxanthin with lard is possibly linked to the increased content of 16:0, 18:1n − 9 or 18:2n − 6 fatty acids in this diet, or a reduction in very long chain monoenes (20:1n − 9 and 22:1n − 9). This suggests that the solubility of astaxanthin is higher in diets containing higher levels of 16:0 or 18:1n − 1, or alternatively, that reductions in longer chain monoenes (20:1n − 9 and 22:1n − 9) increase the micellar solubility of this pigment.     

Effect of protein source and level on growth and performance of the captive freshwater prawn,Macrobrachium rosenbergii

Growth, feed conversion and survival were determined for juvenile Macrobrachium rosenbergii held in tanks under semi-controlled environmental conditions. Feeding trials incorporated water-stable diets at three levels of protein (15, 25 and 35%). The principal protein source combinations consisted either of soybean and tuna meal, or of soybean, tuna and shrimp meal. In a 244-day comparison of these diets, higher protein content produced larger prawns (P < 0.01), but differences between sources were not significant. No significant differences existed between feed conversion ratios (range 1.36–1.72) or percentage survival (range 90.3–93.6%). Trials of several other diets were also conducted, including soybean and Tilapia meal, and copra and Tilapia meal (25% protein level) as principal protein source combinations. After 167 days on these diets, growth was inferior to that obtained with soybean and tuna meal or soybean, tuna and shrimp meal combinations. No significant differences existed between feed conversion ratios or percentages of survival.For the 244 days, a control group of prawns received no formulated diet. Growth and survival in this group during the first 110 days suggested that naturally occurring algae contributed substantially to the prawns' nutrition.Mean prawn length after 244 days on the best diet (35% protein from soybean and tuna meal) was 73 mm, and growth rate was equivalent to that achieved under pilot pond conditions.