Artikel
Does spinal cord injury influence endogenous neural stem progenitor cell characteristics? In-vivo and in-vitro studies in a rat spinal cord injury model
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Autoren
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F. Knerlich-Lukoschus
- Department of Neurosurgery, University of Schleswig-Holstein Medical Center, Kiel, Germany; Department of Anatomy, University of Kiel, Germany
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S. Krossa
- Zentrum für Biochemie & Molekularbiologie, University of Kiel, Germany
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R. Lucius
- Department of Anatomy, University of Kiel, Germany
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H.M. Mehdorn
- Department of Neurosurgery, University of Schleswig-Holstein Medical Center, Kiel, Germany
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J. Held-Feindt
- Department of Neurosurgery, University of Schleswig-Holstein Medical Center, Kiel, Germany
| Veröffentlicht: | 28. April 2011 |
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Gliederung
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Objective: Recruitment of endogenous self-renewing neural stem progenitor cells (NSPCs) provides a powerful tool for spinal cord injury (SCI) repair strategies. The characterization of the NSPC properties is essential for creating such innovative approaches.
Methods: Spinal cord (sc) lesions (200kdyn; Infinite-Horizon-Impactor) were applied at T8 in adult male Long Evans rats. Sham controls received laminectomies. (1) Detection of NSPC in vivo: cell proliferation was studied via BRDU labeling at different post surgery time points in combination with double labeling (stem cells: Nestin, Musashi; major cells: ß-III tubulin, CNPase, GFAP). (2) To further characterize sc NSPC in vitro, NSPCs were isolated from thoracic sc (lesion vs. sham) and cultured according to established protocols. Cell proliferation was determined (CyQuant) and differentiation capacities were examined (real-time RT PCR, Immunohistochemistry). (3) To evaluate influences of microenvironmental changes on NSPCs differentiation, NSPCs were cultured in the presence and absence of chemokines (CCL2, CCL3).
Results: (1) After lesion application, BRDU-labeled cells were detectible on the lesion level (peak 48 h to 7 d). These cells co-labeled with stem cell markers and chemokines. (2) Isolated NSPCs exhibited neurosphere morphology and Nestin/Musashi labeling. Cell proliferation after sham and lesion were similar. NSPC could be differentiated into three major cells lines (astroglia, oligodendroglia, neurons): after lesion application, there were less major oligodendrocytes detectible compared to the sham, but more GFAP and ß-III tubulin + cells. (3) Preliminary data on the influence of chemokine on NSPC showed an effect of CC chemokines in that there was more differentiation into GFAP + cells.
Conclusions: SCI induce NSPC proliferation in vivo, which was partially co-labeled with chemokines. Isolated NSPC behave differently after lesion application in that they differentiate less into major oligodendrocytes as after sham operation. Chemokines influence differentiation directions, thus providing therapeutic targets for SCI.
