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Open Access Highly Accessed Research article

Isolation of mineralizing Nestin+ Nkx6.1+ vascular muscular cells from the adult human spinal cord

Daria Mamaeva1, Chantal Ripoll2, Claire Bony3, Marisa Teigell2, Florence E Perrin45, Bernard Rothhut2, Ivan Bieche6, Rosette Lidereau6, Alain Privat2, Valérie Rigau7, Hélène Guillon2, Florence Vachiery-Lahaye7, Daniele Noel3, Luc Bauchet7 and Jean-Philippe Hugnot28*

Author Affiliations

1 CNRS UPR 1142, Institute of Human Genetics, 141, rue de la Cardonille, 34396 Montpellier Cedex 05, France

2 INSERM U1051, Institut des Neurosciences de Montpellier, Hôpital St ELOI, BP 74103 80, av Augustin Fliche, 34091 Montpellier Cedex 05, France

3 INSERM U844, Institut des Neurosciences de Montpellier, Hôpital St ELOI, BP 74103 80, av Augustin Fliche, 34091 Montpellier Cedex 05, France

4 IKERBASQUE, Basque Foundation for Science, 48011 Bilbao, Spain

5 Department of Neuroscience, Faculty of Medicine and Odontology, University of the Basque Country UPV/EHU, 48940 Leioa, Spain

6 INSERM U735, Institut Curie - Hôpital René Huguenin, 35 rue Dailly, 92210 Saint-Cloud, France

7 CHU Montpellier, Hopital Guy de Chaulliac, Montpellier, France

8 Université Montpellier 2, Place Eugène Bataillon, 34095 Montpellier cedex 5, France

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BMC Neuroscience 2011, 12:99  doi:10.1186/1471-2202-12-99

Published: 10 October 2011

Abstract

Background

The adult central nervous system (CNS) contains different populations of immature cells that could possibly be used to repair brain and spinal cord lesions. The diversity and the properties of these cells in the human adult CNS remain to be fully explored. We previously isolated Nestin+ Sox2+ neural multipotential cells from the adult human spinal cord using the neurosphere method (i.e. non adherent conditions and defined medium).

Results

Here we report the isolation and long term propagation of another population of Nestin+ cells from this tissue using adherent culture conditions and serum. QPCR and immunofluorescence indicated that these cells had mesenchymal features as evidenced by the expression of Snai2 and Twist1 and lack of expression of neural markers such as Sox2, Olig2 or GFAP. Indeed, these cells expressed markers typical of smooth muscle vascular cells such as Calponin, Caldesmone and Acta2 (Smooth muscle actin). These cells could not differentiate into chondrocytes, adipocytes, neuronal and glial cells, however they readily mineralized when placed in osteogenic conditions. Further characterization allowed us to identify the Nkx6.1 transcription factor as a marker for these cells. Nkx6.1 was expressed in vivo by CNS vascular muscular cells located in the parenchyma and the meninges.

Conclusion

Smooth muscle cells expressing Nestin and Nkx6.1 is the main cell population derived from culturing human spinal cord cells in adherent conditions with serum. Mineralization of these cells in vitro could represent a valuable model for studying calcifications of CNS vessels which are observed in pathological situations or as part of the normal aging. In addition, long term propagation of these cells will allow the study of their interaction with other CNS cells and their implication in scar formation during spinal cord injury.

Keywords:
human central nervous system; spinal cord; stem cells; vascular muscle cells; osteogenesis; Nkx6.1; calcification