Project description:Red Jungle Fowl (male and female) tissues were analyzed using LC-MS/MS. Tissues analyzed were: adipose, adrenal gland, breast muscle, cerebellum, cerebrum, gonad, heart, hypothalamus, kidney, liver, lung, pancreas, proventriculus, sciatic nerve, speen. Samples were analyzed using an LTQ Velos Pro mass spectrometer. Xtandem was used to perform spectrum searches. Databases included are NCB refseq, Ensembl, and 6-frame translation of the chicken genome.
Project description:Ma-Huang chicken as a high-quality broiler is one of the most popular chickens in the frozen chicken market. However, some chicken may instead have white or lighter skin, which directly causes economic losses every year. To obtain better insight into the molecular mechanisms associated with the process of pigmentation of yellow-skinned broilers reared under intensive conditions, a total of six-hundred Ma-Huang chickens was randomly selected in a single slaughterhouse, the color measurements were carried out on both cloaca(alive) and five different part after slaughtering adopting the L* a* b* system and using a 3nh-NH310 colorimeter, color values from areas of the chicken skin pear each image automatically retrieved by MATLAB, production and slaughtering traits were also measured, comparative transcriptomic analysis of high yellowness(s_deep) versus low yellowness(s_light) skin was performed using the Illumina Hiseq 4000. Average values of the cloaca(alive), cloaca (hair removal), thigh, shank and abdominal fat were 8.98, 7.66, 2.62, 7.29 and 12.86, respectively. The better production and slaughtering traits were observed in higher skin yellowness chicken. Yellowness values of the cloaca(alive) and after slaughtering were significantly correlated (p < 0.05), suggesting that the color of after slaughtering evaluation may be carried out on cloaca(alive). A total of 19061 unigenes were assembled from the reads obtained from the skin of two groups, 882 unigenes were differentially expressed between s_deep and s_light (Fold change ≥ 2, Adjusted P <= 0.001), 612 that up-regulated and 270 that down-regulated genes in s_deep skin, as compared with s_light skin. Twelve promising candidate genes may play an important role in the pigmentation of chicken skin, i.e. GPR143, PMEL, TYR, CYP11A1, TECRL, ACACB, TLR2B, ALDH1A3, FHL2, TECRL, DUOX2, SMOC1 were included. Furthermore, some important functional pathways were revealed, such as the biological process, cellular component and molecular function, which appear to be much activity in skin pigmentation. Our data provide a valuable resource for identifying genes whose functions are critical to skin pigmentation, facilitate understanding the molecular mechanisms of the skin color variation on yellow-skinned broiler chickens under commercial conditions, accelerate the molecular selection of the specific strain on consistent skin colors which allow reduction in pigment use to achieve the skin acceptance by the consumer.
Project description:Cultured skin substitutes, prepared using keratinocytes, fibroblasts and biopolymers, can facilitate closure of massive burn wounds by increasing the availability of autologous tissue for grafting. However, because they contain only two cell types, skin substitutes cannot replace all of the functions of native human skin. To better understand the physiological and molecular differences between cultured skin substitutes and native skin, we undertook a comprehensive analysis of gene expression in native skin, cultured keratinocytes, cultured fibroblasts, and skin substitutes using Affymetrix gene chip microarrays. Goals: Our analysis focused on identifying gene signatures that were highly characteristic of each cell and tissue type, and those that are regulated by the formation of cultured skin substitute from the individual components. Normalization: We used a normalization and referencing strategy that consisted of BioConductor/RMA Express RMA processing of the entire series of cel files followed by a per gene normalization in which the median value of expression for each gene was derived from the cultured samples only, and this was used as a reference for all samples including the cultured skin substitute. This approach allowed for the identification of genes that were higher and lower-expressed in the cultured skin relative to the individual cell types that were also expressed strongly or weakly in normal skin relative to the median value established by the three cell types. Results Summary:We identified six major clusters of coordinately regulated genes that were the most differentially expressed between groups. These clusters correspond to biomarker pools representing expression signatures for native skin, fibroblasts, keratinocytes, and cultured skin. The expression analysis revealed that entire clusters of genes were either up-regulated or down-regulated upon combination of fibroblasts and keratinocytes in cultured skin grafts. Further, several categories of genes were overexpressed in cultured skin substitutes compared with native skin, including genes associated with hyperproliferative skin or activated keratinocytes. The observed pattern of expression indicates that cultured skin substitutes in vitro, which display a well-differentiated epidermal layer, exhibit skin-like differentiation relative to gene expression patterns in the individual cells. This consists of both the activation of normal skin signature genes and the suppression of keratinocyte and fibroblast signatures. There is also a signature consistent with a hyperproliferative phenotype similar to wounded native skin. Keywords: Cell interaction and co-culture response expression profile
Project description:Global demand for animal protein is expected to double by 2050 at a time when resource-intensive livestock production is reaching peak capacity. Cellular agriculture offers an opportunity to meet the increasing demand for animal products, but the technology is currently limited by the genetic stability of immortalized cells, low culture yields, and high production costs. Here we demonstrate the spontaneous immortalization and long-term genetic stability of fibroblasts derived from the indigenous Israeli Baladi and the commercial Broiler Ross chicken breeds. Cells were adapted for growth as single-cell suspensions in animal-component free culture medium reaching cell densities of over 100x10^6 cells/mL in ATF perfusion presenting a production yield of over 33% w/v. We show that soy phosphatidylcholine, a major component of lecithin, activates PPAR, driving adipogenesis in immortalized chicken fibroblasts. Harvesting cultured adipocytes and blending them with high moisture extruded soy protein formed cultured chicken strips in which mouth feel and texture were supported by a blend of animal and plant proteins while aroma and flavor were driven by cultured animal fat. Visual and sensory analysis graded the product 4.5 out of 5.0, with over 85% percent of the study group said they are extremely likely to replace their food choice with this cultured meat product. The ability to create immortalized lines without genetic modification and the high yield process for cultured meat production presents an important steppingstone in the market realization of cultured meat.
Project description:The growth factors and signaling pathways that are essential for the inducing proliferation of chicken PGCs is unclear. We investigated the effect of basic fibroblast growth factor (bFGF) on the survival and proliferation of PGCs under feeder-free conditions. We used microarrays to examine the genes regulated by bFGF treatment in chicken PGCs. Cultured PGCs were collected before and after withdrawal of bFGF for 24 h, and 24 h after bFGF replacement. The RNA was extracted and hybridized on Affymetrix microarrays. Three replicates each.
Project description:Cultured skin substitutes, prepared using keratinocytes, fibroblasts and biopolymers, can facilitate closure of massive burn wounds by increasing the availability of autologous tissue for grafting. However, because they contain only two cell types, skin substitutes cannot replace all of the functions of native human skin. To better understand the physiological and molecular differences between cultured skin substitutes and native skin, we undertook a comprehensive analysis of gene expression in native skin, cultured keratinocytes, cultured fibroblasts, and skin substitutes using Affymetrix gene chip microarrays. Goals: Our analysis focused on identifying gene signatures that were highly characteristic of each cell and tissue type, and those that are regulated by the formation of cultured skin substitute from the individual components. Normalization: We used a normalization and referencing strategy that consisted of BioConductor/RMA Express RMA processing of the entire series of cel files followed by a per gene normalization in which the median value of expression for each gene was derived from the cultured samples only, and this was used as a reference for all samples including the cultured skin substitute. This approach allowed for the identification of genes that were higher and lower-expressed in the cultured skin relative to the individual cell types that were also expressed strongly or weakly in normal skin relative to the median value established by the three cell types. Results Summary:We identified six major clusters of coordinately regulated genes that were the most differentially expressed between groups. These clusters correspond to biomarker pools representing expression signatures for native skin, fibroblasts, keratinocytes, and cultured skin. The expression analysis revealed that entire clusters of genes were either up-regulated or down-regulated upon combination of fibroblasts and keratinocytes in cultured skin grafts. Further, several categories of genes were overexpressed in cultured skin substitutes compared with native skin, including genes associated with hyperproliferative skin or activated keratinocytes. The observed pattern of expression indicates that cultured skin substitutes in vitro, which display a well-differentiated epidermal layer, exhibit skin-like differentiation relative to gene expression patterns in the individual cells. This consists of both the activation of normal skin signature genes and the suppression of keratinocyte and fibroblast signatures. There is also a signature consistent with a hyperproliferative phenotype similar to wounded native skin. Experiment Overall Design: The sample series consists of native human skin (NHS) samples isolated from female donors undergoing reduction mammoplasty (breast skin) or abdominoplasty (abdomen skin). Skin samples from donors that were used to establish cultures of fibroblasts (CF) and keratinocytes (CK) were assigned donor numbers in the order they were processed in the laboratory, for example: 633, 634, etc. An additional human skin sample (C-1-Ref) was used only to make RNA as a standard control, and was therefore not assigned a donor number. Cultured skin substitutes (CSS) were prepared using isogenic CF and CK from each donor, and were cultured for 2 weeks in vitro to permit development of a stratified and cornified epidermal layer (confirmed by histology). For microarray analysis, RNA was isolated from intact NHS, from CF and CK in monolayer cultures, and from CSS. Samples are labeled indicating the sample type and donor number; for example, CF633 represents cultured fibroblasts from donor 633. To control for variation between individuals, four donors (= biological replicates) were used for each sample type: NHS, CF, CK, and CSS. Efforts were made to have complete sets of 4 samples from each donor, but intact RNA was not obtainable from 2 of the NHS samples (donors 634 and 651); these were replaced with NHS RNA from similar donors (donors C-1-Ref and 636). To check the fidelity of the microarray analysis, 2 of the RNA samples (CK639 and CSS651) were analyzed in duplicate (= technical replicates)
Project description:Global demand for animal protein is expected to double by 2050 at a time when resource-intensive livestock production is reaching peak capacity. Cellular agriculture offers an opportunity to meet the increasing demand for animal products, but the technology is currently limited by the genetic stability of immortalized cells, low culture yields, and high production costs. Here we demonstrate the spontaneous immortalization and long-term genetic stability of fibroblasts derived from the indigenous Israeli Baladi and the commercial Broiler Ross chicken breeds. Cells were adapted for growth as single-cell suspensions in animal-component free culture medium reaching cell densities of over 100x10^6 cells/mL in ATF perfusion presenting a production yield of over 33% w/v. We show that soy phosphatidylcholine, a major component of lecithin, activates PPAR, driving adipogenesis in immortalized chicken fibroblasts. Harvesting cultured adipocytes and blending them with high moisture extruded soy protein formed cultured chicken strips in which mouth feel and texture were supported by a blend of animal and plant proteins while aroma and flavor were driven by cultured animal fat. Visual and sensory analysis graded the product 4.5 out of 5.0, with over 85% percent of the study group said they are extremely likely to replace their food choice with this cultured meat product. The ability to create immortalized lines without genetic modification and the high yield process for cultured meat production presents an important steppingstone in the market realization of cultured meat.
Project description:To investigate whether skin bacteria might influence the expression of selected genes, we co-cultured human keratinocytes with S. epidermidis, an abundant commensal in human skin and performed RNA sequencing analysis.
Project description:The dorsal and leg skin showed significant difference in skin thickness, feather follicle density and size.These traits are important characters of slaughtered chicken appearance. Global gene expression profiling was conducted in dorsal and leg skin of chickens, using the Agilent Chicken Gene Expression Chip. A total of 676 differentially expressed genes (DEGs) with at least 2-fold differences were identified (P < 0.05).GO analysis showed that DEGs were significantly involved in cell proliferation, differentiation, apoptotic, cell-cell adhesion and Wnt signaling pathway; KEGG pathway analyses found that DEGs were significantly mapped into ECM-receptor interaction, focal adhesion, Cell adhesion molecules (CAMs), regulation of actin cytoskeleton and Hedgehog signaling pathway.