An Improved Harvest and in Vitro Expansion Protocol for Murine Bone Marrow-Derived Mesenchymal Stem Cells
Joint Authors
Van Riet, Ivan
Xu, Song
Van Camp, Ben
Vanderkerken, Karin
De Becker, Ann
Source
Issue
Vol. 2010, Issue 2010 (31 Dec. 2009), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-10-27
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
Compared to bone marrow (BM) derived mesenchymal stem cells (MSCs) from human origin or from other species, the in vitro expansion and purification of murine MSCs (mMSCs) is much more difficult because of the low MSC yield and the unwanted growth of non-MSCs in the in vitro expansion cultures.
We describe a modified protocol to isolate and expand murine BM derived MSCs based on the combination of mechanical crushing and collagenase digestion at the moment of harvest, followed by an immunodepletion step using microbeads coated with CD11b, CD45 and CD34 antibodies.
The number of isolated mMSCs as estimated by colony forming unit-fibroblast (CFU-F) assay showed that this modified isolation method could yield 70.0% more primary colonies.
After immunodepletion, a homogenous mMSC population could already be obtained after two passages.
Immunodepleted mMSCs (ID-mMSCs) are uniformly positive for stem cell antigen-1 (Sca-1), CD90, CD105 and CD73 cell surface markers, but negative for the hematopoietic surface markers CD14, CD34 and CD45.
Moreover the immunodepleted cell population exhibits more differentiation potential into adipogenic, osteogenic and chondrogenic lineages.
Our data illustrate the development of an efficient and reliable expansion protocol increasing the yield and purity of mMSCs and reducing the overall expansion time.
American Psychological Association (APA)
Xu, Song& De Becker, Ann& Van Camp, Ben& Vanderkerken, Karin& Van Riet, Ivan. 2010. An Improved Harvest and in Vitro Expansion Protocol for Murine Bone Marrow-Derived Mesenchymal Stem Cells. BioMed Research International،Vol. 2010, no. 2010, pp.1-10.
https://search.emarefa.net/detail/BIM-988737
Modern Language Association (MLA)
Xu, Song…[et al.]. An Improved Harvest and in Vitro Expansion Protocol for Murine Bone Marrow-Derived Mesenchymal Stem Cells. BioMed Research International No. 2010 (Dec. 2010), pp.1-10.
https://search.emarefa.net/detail/BIM-988737
American Medical Association (AMA)
Xu, Song& De Becker, Ann& Van Camp, Ben& Vanderkerken, Karin& Van Riet, Ivan. An Improved Harvest and in Vitro Expansion Protocol for Murine Bone Marrow-Derived Mesenchymal Stem Cells. BioMed Research International. 2010. Vol. 2010, no. 2010, pp.1-10.
https://search.emarefa.net/detail/BIM-988737
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-988737