[Frontiers in Bioscience 14, 4281-4298, January 1, 2009]

Methodology, biology and clinical applications of mesenchymal stem cells

Lindolfo da Silva Meirelles1, Nance Beyer Nardi2

1Center for Cell Therapy, Centro Regional de Hemoterapia de Ribeirao Preto - HCFMRP, Universidade de Sao Paulo, Rua Tenente Catao Roxo 2501, 14051-140, Ribeirao Preto, SP, Brazil, 2Departament of Genetics, Universidade Federal do Rio Grande do Sul, Av. Bento Goncalves 9500, 91501-970, Porto Alegre, RS, Brazil

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Conditions employed for MSC isolation, culture expansion, and differentiation
4. MSCs found in several different organs/tissues
5. MSC differentiation potential and plasticity
6. MSC identity traced to pericytes in vivo
7. MSCs as trophic mediators
8. Immunoregulatory function of mesenchymal stem cells
9. Perivascular stem cells and tissue regeneration
10. Clinical and pre-clinical applications of MSCs
10.1. Myocardial infarction
10.2. Fibrosis
10.3. Immunological diseases
10.4. Stroke
10.5. Angiogenesis and cancer therapy
11. Methods for the in vitro study of mesenchymal stem cells
11.1. Isolation and culture of murine bone marrow MSCs
11.1.1. Reagents
11.1.2. Procedure
11.1.3. Observations
11.2. Isolation and culture of MSCs from other murine organs and tissues
11.2.1. Procedure
11.3. Isolation and culture of MSCs from human bone marrow
11.3.1. Additional reagents
11.3.2. Procedure
11.4. Isolation and culture of MSCs from human liposuction material
11.4.1. Additional reagents
11.4.2. Procedure
11.5. CFU-F assay
11.5.1. Procedure
11.6. Differentiation assays
11.6.1. Osteogenic differentiation
11.6.1.1. Additional reagents
11.6.1.2. Procedure
11.6.2. Adipogenic differentiation
11.6.2.1. Additional reagents
11.6.2.2. Procedure
11.6.3. Chondrogenic differentiation
11.6.3.1. Additional reagents
11.6.3.2. Procedure
12. Conclusion
13. Acknowledgements
14. References

1. ABSTRACT

Mesenchymal stem cells (MSCs) are adult stem cells able to give rise to mature mesenchymal cell types. Plastic-adherent cells are operationally defined as MSCs based on their ability to proliferate and differentiate into cells such as osteoblasts, adipocytes and chondrocytes. In the past ten years, cultured MSCs have been shown to exhibit great plasticity in culture, as they can differentiate into cells with ectodermal and endodermal characteristics, suggesting their use as a source of cells to treat different diseases. More recently, cultured MSCs were found to secrete various bioactive molecules that display anti-apoptotic, immunomodulatory, angiogenic, anti-scarring, and chemoattractant properties, providing a basis for their use as tools to create local regenerative environments in vivo. Whereas the properties of cultured MSCs have been studied for a long time, their exact location in vivo is slowly becoming apparent as evidence indicates that pericytes behave as stem cells throughout the organism. In this review, we discuss some aspects of MSC basic biology, the methodology involved in MSC culture, and some clinical and pre-clinical applications of cultured MSCs.