Comparative growth kinetics and functional gene expression analysis in different media formulations for porcine mesenchymal stem cells in vitro

2^nd Indo-Global Summit & Expo on Veterinary

October 26-28, 2015 Hyderabad, India

Sangeetha Kannan1, Shree Vidhya Srinivasan1, Punith B D1, Sujoy K Dhara2 and Jyotirmoy Ghosh1

1National Institute of Animal Nutrition and Physiology, India 2Indian Veterinary Research Institute, India

Scientific Tracks Abstracts: J Veterinar Sci Technol

Abstract :

Porcine mesenchymal stem cells (pMSC) are useful in human regenerative therapies and transgenic animal production. However, the conventional basal media formulation is not best suited for long term in vitro expansion of these cells. The study was thus designed to test the suitability of three commonly used basal media formulations, namely alpha minimum essential media(�±MEM), advanced Dulbeccoâ��s Modified Eagle Medium (aDMEM), tissue culture media 199 (M199) alone and their 1:1 combinations (�±MEM/ M199, aDMEM/M199, aDMEM/�±MEM) with the same standard culture supplements for derivation and propagation of bone marrow pMSC. The pMSC derived and grown from the same animals in different basal media were tested for the growth kinetics and expression status of cell proliferation, osteogenic, adipogenic and chondrogenic differentiation marker genes viz., cyclin dependant kinase II (CDK-II), osteocalcin, CCAAT-enhancer binding protein �± (CEBP �±) and Collagen2 (Col2), respectively at passage 5. The growth kinetics based on the population doubling showed the �±MEM/M199 media had the least doubling time of 56.3�±10.18 hours compared to the cells derived and grown in any other basal media formulations alone or in combination (above 80 hours). Coinciding with the proliferation rate the expression of cell proliferation marker CDK-II gene was found maximum in �±MEM/M199 combinations as compared to other media formulations. The lower expression of differentiation marker genes, osteocalcin, CEBP and Col2 in �±MEM/M199 revealed that thecells in this media remained more undifferentiated compared to other media formulations. The results of this study indicated that the �±MEM/M199 combination might be a better basal media for derivation and culture of bone marrow derived pMSC.

Biography :

Sangeetha Kannan has completed her Master’s degree from PESIT, Bangalore University in 2006. She has then worked as DST-Woman Scientist A project under the Guidance of Dr. Jyotirmoy Ghosh, Senior Scientist, NIANP, Bangalore. She is also pursuing her PhD under Jain University, Bangalore.

Email: sangeetha_k26@rediffmail.com