Reprogramming of buffalo (Bubalus bubalis) foetal fibroblasts with avian egg extract for generation of pluripotent stem cells

An attempt was made to study the efficiency of chicken (Gallus gallus domesticus) egg extract (EE) to reprogram buffalo (Bubalus bubalis) foetal fibroblasts (bFFs) without incorporation of ectopic transcription factors. The isolated bFFs were cultured in media supplemented with 2%, 4%, 6% and 10% EE...

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Bibliographic Details
Published in:Research in veterinary science 2014-04, Vol.96 (2), p.292-298
Main Authors: Mahapatra, Puspendra S., Bag, Sadhan
Format: Article
Language:English
Subjects:
Alkaline Phosphatase - analysis
Animals
Avian
Biomedical research
Buffalo
Buffaloes - genetics
Buffaloes - physiology
Chickens
Egg extract
Eggs
Female
Fetus
Fibroblasts - cytology
Foetal fibroblast
Gene expression
Genomes
Homeodomain Proteins - genetics
Induced Pluripotent Stem Cells - cytology
Lewis X Antigen - genetics
Methods
Octamer Transcription Factor-3 - genetics
Pregnancy
Proteins
Reprogramming
Reverse Transcriptase Polymerase Chain Reaction - veterinary
RNA - chemistry
RNA - genetics
Stem cells
Transcription factors
Veterinary medicine
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Summary:An attempt was made to study the efficiency of chicken (Gallus gallus domesticus) egg extract (EE) to reprogram buffalo (Bubalus bubalis) foetal fibroblasts (bFFs) without incorporation of ectopic transcription factors. The isolated bFFs were cultured in media supplemented with 2%, 4%, 6% and 10% EE to induce reprogramming. It was observed that fewer but larger sized alkaline phosphatase positive (AP+ve) colonies developed in culture system containing 2% EE whereas, more but smaller sized colonies developed in 4%, 6% and 10% EE. The developed colonies expressed pluripotency markers like Oct4, Nanog, SSEA1, TRA-1-60, TRA-1-81 and RT-PCR study revealed relative expression of genes indicating pluripotency (Oct4, Sox2, Nanog and FoxD3) increased as the concentration of EE increased in culture systems confirming the reprogramming capability of chicken EE.
ISSN:0034-5288
1532-2661
DOI:10.1016/j.rvsc.2014.02.008