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Arch Iran Med. 2018;21(8): 362-367.
PMID: 30113858
Scopus ID: 85057056609
  Abstract View: 3367
  PDF Download: 1847

Original Article

Use of Some Additives for Improving Mesenchymal Stem Cell Isolation Outcomes in Non-Mobilized Peripheral Blood

Noushin Pouryazdanpanah 1, Reza Vahidi 2,3, Shahriar Dabiri 4, Ali Derakhshani 2,4*, Alireza Farsinezhad 1,3*

1 1 Department of Hematology and Laboratory Sciences, Faculty of Allied Medical Sciences, Kerman University of Medical Sciences, Kerman, Iran
2 Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, Kerman, Iran
3 Cell Therapy and Regenerative Medicine Comprehensive Center, Kerman University of Medical Sciences, Kerman, Iran
4 Pathology and Stem Cell Research Center, Kerman University of Medical Sciences, Kerman, Iran
*Corresponding Authors: *Corresponding Authors: Ali Derakhshani, PhD; Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, Kerman, Iran. Tel/Fax: +98- 34-33257318, Email: Derakhshani.sci@gmail.com; Alireza Farsinejad, PhD; Cell Therapy and Regenerative Medicine Comprehensive Center, Kerman University of Medical Sciences, Kerman, Iran . Tel/Fax: +98- 34-33257318, Email: , Email: farsinezhad239@yahoo.com; Email: farsinezhad239@yahoo.com

Abstract

Background: The mesenchymal stem cells (MSCs) of peripheral blood (PB) have been recognized as a promising source for allogeneic cell therapy. The objective of the present study was to isolate and characterize MSCs derived from non-mobilized PB, and evaluate their differentiation potential.

Methods: The buffy coat mononuclear fractions of the PB were concentrated using the Ficoll-Paque density gradient centrifugation and were grown on primary and secondary culture media, respectively. The isolated cells were characterized using a multidisciplinary approach which was based on morphology, immunophenotyping, gene expression, and multipotentiality. Flow cytometry and Reverse transcription polymerase chain reaction (RT-PCR) were used to identify the expression of different MSC markers. Finally, after culturing in osteogenic and adipogenic induction media, the isolated cells were stained by Alizarin red and Oil-Red O.

Results: In spite of absence of any bone marrow stimulating factor, the isolation approach in this study yielded a rather homogeneous and spindle-shaped mononuclear cell population (the yield of passage 0 was 0.65 ± 0.15) that stained positive for CD90, CD105, and CD73, and were negative for CD45 and CD34. These cells have high proliferative capacity (confirmed by the expression of Oct-4, Nucleostemin, and Nanog genes) and were able to differentiate into lineage-specific committed cells, when exposed to the appropriate medium.

Conclusions: Overall, it can be concluded that conventional, labour-intensive and time-consuming approaches are not necessary in isolating MSCs from PB. This relatively accessible and minimally invasive source, PB, represents a good alternative reservoir of homogeneous MSCs that could open a new era for practical exploitation in regenerative medicine.


Cite this article as: Pouryazdanpanah N, Vahidi R, Dabiri S, Derakhshani A, Farsinejad AR. Use of some additives for improving mesenchymal stem cell isolation outcomes in non-mobilized peripheral blood. Arch Iran Med. 2018;21(8):362–367.
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Submitted: 10 Dec 2017
Accepted: 12 May 2018
ePublished: 01 Aug 2018
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