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Asian Journal of Animal and Veterinary Advances
Year: 2015  |  Volume: 10  |  Issue: 10  |  Page No.: 537 - 548

Isolation, Culture and Characterization of New Zealand White Rabbit Mesenchymal Stem Cells Derived from Bone Marrow

Mudasir Bashir Gugjoo, Amarpal, Prakash Kinjavdekar, Hari Prasad Aithal, Mohd Matin Ansari, Abhijit MotiRam Pawde and Gutulla Taru Sharma    

Abstract: Mesenchymal stem cells are recognised based upon the plastic adherence, fibroblastic morphology, expression of certain surface markers, non-expression of haematopoetic markers and their ability to differentiate into atleast three lineages viz., adipogenic, chondrogenic and osteogenic. The rabbit Mesenchymal Stem Cells (rMSCs) though used extensively in research but have not been thoroughly studied and are not compared to other species. The present study was therefore conducted to determine the morphology, surface markers and trilineage differentiation potential of New Zealand white rabbit MSCs. Isolation of rMSCs was done by an established method of density gradient method using Ficoll-hapaque. The cells were characterised by Phase Contrast Microscopy, Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Alkaline Phosphatase (AP) staining. The cells isolated were plastic adherent and had fibroblastic spindle shape with eccentric irregular nuclei. The cells expressed surface markers viz., CD 105 and CD 106 besides expressing genes of collagen type II and I. Haematopoetic markers (CD 34 and CD 45) and aggrecan gene however, were not expressed. The rMSCs showed a moderate alkaline phosphates activity. Trilineage differentiation was conducted utilizing prepared differentiation media and rMSCs were differentiated into corresponding cell lineages based upon the medium used. It was concluded that rMSCs possess morphology similar to other species with good proliferation rate and exhibit the characteristics laid down by International Society for Cellular Therapy (ISCT). The present study provided basic protocols for characterization of rabbit MSCs that should be used before application of these cells for any research or therapy.

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