Study on Cloning, Sequencing and Phylogenetic Analysis of Granulocyte Macrophage Colony Stimulating Factor of Indian Cattle (Bos indicus)

The cDNA coding for Granulocyte macrophage colony stimulating factor (GM-CSF) from LPS activated macrophages was amplified using reverse transcriptase polymerase chain reaction (RT-PCR), which was then cloned into the pCNDA vector and sequenced. The amplified bovine GM-CSF cDNA was 755 bp length without the secretory signal peptide sequence. The ORF coded for 143 amino acids with a molecular weight of 16200 Da and was 99 percent identical to Bos taurus and Bubalus bubalis, respectively. The phylogenetic study found that the GM-CSF sequences of swine and horse are the most distinct from those of buffalo, sheep, goat, and deer.

Author (S) Details

Ramya Kalaivanan

Veterinary College and Research Institute, Namakkal-02, India. and Indian Veterinary Research Institute, Hebbal, India.

P. Sankar

Veterinary College and Research Institute, Namakkal-02, India.

Subodh Kishore

Indian Veterinary Research Institute, Hebbal, India.

View Book :- https://stm.bookpi.org/RAAVS-V2/article/view/4078

Optimizing in vitro Generation of Interleukin-10 Secreting B Regulatory Cells

Bregulatory cells are a subpopulation of IL-10-secreting B-cells that are frequently identified as CD19+/38hi/24hi (B-regs). Changes in stimulating molecule concentration have an influence on the population of human Interleukin-10 secreting B-regs. For possible cell treatment, B-regs can be generated by in-vitro co-culture of adipose tissue derived mesenchymal stromal cells (AD-MSC) and peripheral blood mononuclear cells (PBMC). In the co-culture of AD-MSC and peripheral blood mononuclear cells (LPS-EK), lipopolysaccharide–E.coli–K12 strain (LPS-EK) was added in various doses. cellular (PBMC). The concentration of IL-10 in the cell supernatant was measured using a quantitative ELISA at each time point. Cells were examined for morphology, sterility, total count, vitality, and immune-phenotype. The data was evaluated to establish the time interval during which the greatest B-reg population and IL-10 secretion were achieved. The current study adds to our understanding of the kinetics of IL-10 secretion by human B-regs derived from AD-MSC and PBMC in vitro. The current research focuses on the in vitro production of B-regs from mesenchymal stem cells generated from adipose tissue. stromal cells (AD-MSC) and peripheral blood mononuclear cells (PBMC), as well as a partial characterisation of B-regs in terms of IL-10 secretion, time, and the quantity of stimulating molecule (i.e. LPS-EK) required for optimal IL-10 secretion. B-regs have been found to secrete cytokine in trace amounts. The media used to feed the cells dilutes the released cytokines by several orders of magnitude. The samples in our investigation were properly concentrated so that the cytokines could be detected using a quantitative Enzyme Linked Immunosorbent Assay (ELISA).

Author (s) Details

K. S. Gupte
Department of Cell Therapy and Regenerative Medicine. G.R. Doshi and K.M. Mehta Institute of Kidney Diseases & Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), India.

A. V. Vanikar
Department of Cell Therapy and Regenerative Medicine. G.R. Doshi and K.M. Mehta Institute of Kidney Diseases & Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), India and Department of Pathology, Laboratory Medicine, Transfusion Services and Immunohematology. G.R. Doshi and K.M. Mehta Institute of Kidney Diseases & ResearchCentre (IKDRC) – Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS),India.

C. N. Patel
Department of Cell Therapy and Regenerative Medicine. G.R. Doshi and K.M. Mehta Institute of Kidney Diseases & Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), India.

U. G. Thakkar
Department of Cell Therapy and Regenerative Medicine. G.R. Doshi and K.M. Mehta Institute of Kidney Diseases & Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), India.

View Book :- https://stm.bookpi.org/RRAB-V10/article/view/2131