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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2018: 95.39
For more details click here

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Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2020.9(2): 967-975
DOI: https://doi.org/10.20546/ijcmas.2020.902.113


Resazurin Based Colorimetric Proliferation Assay for PBMCS of Goats and Sheep
Soumajit Sarkar*, Ashwini Rameshrao Chaple, Aruna Kuniyal, Dhanavelu Muthuchelvan, Muthannan Andavar Ramakrishnan*
Division of Virology, ICAR-Indian Veterinary Research Institute, Mukteswar, Uttarakhand – 263 138, India,
*Corresponding author
Abstract:

Resazurin dye-based colorimetric assay is simple and cost-effective to evaluate viability and cytotoxicity. This dye also used for evaluating proliferative responses of PBMCs against mitogens and antigens especially to assess the CMI response against vaccines and pathogens. However, the assay needs to be optimized for different cell types and animal species. So far, resazurin based assay is not available for sheep and goats. In the present study, the assay was optimized for PBMCs of goat with two density gradients viz., Histopaque 1.077 and 1.083, and different culture conditions. The cell yield obtained through Histopaque 1.083 was 10.5 times more than cells obtained through 1.077. Out of three tested media, cells cultivated with TCM-199 showed the highest growth followed by RPMI-1640, and AIM-V. The resazurin dye @600 µM and 24 h dye reduction time yielded a better result. Among four mitogens tested PHA-P induced higher proliferation followed by ConA, PWM, and LPS. The result was correlated with the result obtained in the chemiluminescence based BrdU ELISA. Further, the resazurin assay also validated in sheep PBMCs.


Keywords: Cell proliferation; goat; sheep; PBMCs; Resazurin; BrdU
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How to cite this article:

Soumajit Sarkar, Ashwini Rameshrao Chaple, Aruna Kuniyal, Dhanavelu Muthuchelvan, Muthannan Andavar Ramakrishnan. 2020. Resazurin Based Colorimetric Proliferation Assay for PBMCS of Goats and Sheep.Int.J.Curr.Microbiol.App.Sci. 9(2): 967-975. doi: https://doi.org/10.20546/ijcmas.2020.902.113