Valproic acid assisted reprogramming of fibroblasts for generation of pluripotent stem cells in buffalo (Bubalus bubalis).

Generation of pluripotent stem cells by reprogramming somatic cells of quality animals has numerous potential applications in agricultural and biomedical sciences. Unfortunately, till now, reprogramming of buffalo fetal fibroblast cells (bFFs) has been very ineffient despite intensive efforts. Here, we attempted to enhance reprogramming efficiency by using the HDAC inhibitor valproic acid (VPA) in bFFs transfected with pLentG-KOSM pseudo virus carrying mouse specific pluripotent genes. FACS analysis revealed that VPA treatment significantly increased (p < 0.05) GFP+ cells in comparison to VPA untreated control. Further, among different concentrations, 1.5 mM VPA was found to be optimal, increasing about 5 fold GFP+ cells and 2.5-fold GFP+ colonies with significantly (P < 0.05) larger size as compared to control. These colonies were further propagated and characterised. The colonies displayed embryonic stem cell (ESC)-like morphology, normal karyotype, and were positive for alkaline phosphatase staining as well as immune-positive for the ESC specific markers Oct4, Nanog, SSEA1, TRA-1-60 and TRA-1-81. The primary colonies revealed significantly higher (P < 0.05) expression of pluripotent genes than control, which declined gradually on subsequent passages. The reprogrammed cells readily formed embryoid bodies in vitro and cells of all three germ layers. These results indicated that VPA treatment of viral transducted cells can improve the generation of induced pluripotent stem cells and help their long term maintenance in buffalo.

In ovo trace element supplementation enhances expression of growth genes in embryo and immune genes in post-hatch broiler chickens.

BACKGROUND: Differential expression of growth- and immunity-related genes and post-hatch performances were evaluated in in ovo zinc (Zn), iodine (I) or selenium (Se) supplemented chicken embryos. RESULT: There was about 9-18% reduction in hatchability of Zn, I or Se supplemented eggs. In ovo trace element supplementation did not improve post-hatch growth. Two-way analysis of data revealed significant effect (P > 0.01) of period, trace elements and their interactions. Expression of hepatic somatotropin, insulin-like growth factor-II and mucin gene was highest at 20(th) embryonic day but decreased during post-hatch periods. In ovo Zn or I supplemented embryos had higher expression of growth-related genes compared to the Se or un-injected control group. Expression of interleukin-6 was higher (P < 0.01) in in ovo I supplemented chicks (2.5-fold) but lower in the Zn and Se groups than in the un-injected control group. However, Zn and Se supplemented chicks had higher cellular immune gene expression. In vivo response to mitogen phytohaemaglutinin was also higher (P < 0.01) in Zn or Se supplemented chicks CONCLUSION: In ovo supplementation of Zn, I and Se did not improve the post-hatch growth, but increased growth-related gene expression. Iodine improved humoral immune gene expression whereas Zn and Se enhanced cell-mediated immune gene expression in broiler chickens. © 2015 Society of Chemical Industry.

In Ovo Administration of Silver Nanoparticles and/or Amino Acids Influence Metabolism and Immune Gene Expression in Chicken Embryos.

Due to their physicochemical and biological properties, silver nanoparticles (NanoAg) have a wide range of applications. In the present study, their roles as a carrier of nutrients and an immunomodulator were tested in chicken embryos. Cysteine (Cys)+NanoAg injected embryos had smaller livers but heavier breasts on the 19th day of embryogenesis. Cys injected embryos had lower oxygen consumption compared to threonine (Thr) or NanoAg injected embryos. The energy expenditure in Thr+NanoAg, or NanoAg injected embryos was higher than Cys or Cys+NanoAg but was not different from uninjected control embryos. Relative expression of the hepatic insulin-like growth factor-I (IGF-I) gene was higher in Cys or NanoAg injected embryos after lipopolysaccharide (LPS) induction. The gene expression of hepatic tumour necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) did not differ among amino acids, NanoAg and uninjected controls in the non-LPS groups, but increased by many folds in the LPS treated NanoAg, Cys and Cys+NanoAg groups. In LPS treated spleens, TNF-α expression was also up-regulated by NanoAg, amino acids and their combinations, but interleukin-10 (IL-10) expression was down-regulated in Thr, Cys or Thr+NanoAg injected embryos. Toll like receptor-2 (TLR2) expression did not differ in NanoAg or amino acids injected embryos; however, toll like receptor-4 (TLR4) expression was higher in all treated embryos, except for Cys+NanoAg, than in uninjected control embryos. We concluded that NanoAg either alone or in combination with amino acids did not affect embryonic growth but improved immunocompetence, indicating that NanoAg and amino acid complexes can act as potential agents for the enhancement of innate and adaptive immunity in chicken.

Prospects of in ovo feeding and nutrient supplementation for poultry: the science and commercial applications--a review.

In ovo supplementation of poultry embryos was first reported several decades ago, but it is only recently that concerted research has been directed at developing the technology for this process to be routinely used by the poultry industry. Although the technology of in ovo feeding was patented more than 10 years ago, it has not been widely adopted by the poultry industry. This review examines the early development of the enteric system of the poultry embryo; defines and distinguishes between in ovo feeding and in ovo nutrient administration; highlights the importance of early feeding of the chick; and discusses the development of in ovo feeding technology and its effects on hatchability, growth, gut health and immune response of chicks. The range of possible nutrients that can be administered is also explored. The limitations associated with embryo development and nutrient metabolism are highlighted, leading to the prediction of the future role of in ovo feeding in the poultry industry.

Modulation of post-hatch growth and immunity through in ovo supplemented nutrients in broiler chickens.

BACKGROUND: Early post-hatch growth and immunity were assessed through in ovo supplementation of nutrients: amino acids (AA), trace elements (TE), fatty acids and vitamins (FAV) grouped under humoral immunity (HI) or cell-mediated immunity (CMI) on the 18th day of incubation at the broad end of the egg using a 25 mm needle. RESULTS: Hatchability in AA groups was better than TE and FAV groups. CMI groups had better hatchability than HI groups. AA and TE groups had higher chick-to-egg weight ratio (P < 0.01) than the FAV group. At 3 weeks of age, a higher body weight (P < 0.01) was recorded in AA for CMI, TE for HI and FAV for HI groups. FAV-injected chicks had a higher bursa weight at hatch, but TE chicks had higher thymus weight at the 3rd week of age. Humoral immune response was not different in in ovo injected chicks compared to control. CMI was higher (P < 0.01) in AA for CMI, TE for CMI and FAV for CMI or HI nutrient-injected chicks. CONCLUSIONS: In ovo injection of AA for CMI and TE for HI may accelerate growth of broiler chickens. In ovo injection of AA, TE or FAV may modulate CMI in chicks.