Artikel
Optimizing a procedure for primary cell cultures of meningiomas
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Autoren
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Mustafa El-Khatib
- Neurochirurgische Klinik der Medizinischen Einrichtungen der Heinrich Heine Universität Düsseldorf, Germany
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Carolin Tepe
- Neurochirurgische Klinik der Medizinischen Einrichtungen der Heinrich Heine Universität Düsseldorf, Germany
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Birgit Senger
- Neurochirurgische Klinik der Medizinischen Einrichtungen der Heinrich Heine Universität Düsseldorf, Germany
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Hans-Jakob Steiger
- Neurochirurgische Klinik der Medizinischen Einrichtungen der Heinrich Heine Universität Düsseldorf, Germany
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Walter Stummer
- Klinik und Poliklinik für Neurochirurgie des Universitätsklinikum Münster, Germany
| Veröffentlicht: | 16. September 2010 |
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Gliederung
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Objective: In malignant tumors such as glioblastomas, stable cell lines for in vitro studies already exist. In meningiomas there are some immortalized cell lines, but these do not cover all subtypes of meningiomas. Therefore, it is necessary to establish a procedure to implement primary cell cultures in order to study the growth behaviour of the different subtypes of meningiomas.
Methods: Based on the protocol of Hardy et al, we modified several steps to optimise the growth conditions for the culturing of primary meningioma cells. Amongst other factors, we changed the storage temperature, the composition of the medium and the time of the procedure's beginning after resection to determine the best circumstances for primary cell cultures of meningiomas. Beyond this, we investigated whether the growth is influenced by collagen-coated culture bottles or not. At the first passage, we used EMA (endothelial membrane antigen) immunohistochemistry to detect meningioma cells.
Results: Until today we have cultured 28 different meningiomas, 20 WHO I, 6 WHO II and 1 WHO III meningiomas. During the first 15 cases, we frequently changed the conditions to establish the definitive procedure step by step. Four of those 15 meningiomas failed to be cultured. Aside from these of five cell cultures, all remaining 19 cases demonstrated EMA positivity after immunohistochemistry. In all EMA positive cell cultures, further culturing of the cells showed a continuous decrease in positivity of EMA. There was no difference of growth or EMA positivity in relation to the grading of meningiomas. The faster the first passage could be done the more positive was the immunohistochemistry.
Conclusions: We were able to optimize the growth conditions by the modification of Hardy's protocol for culturing of primary meningioma cells. But we could not establish any stable cell lines for time independent implementations of further in vitro studies. The best time for in vitro studies in primary meningioma cell cultures seems to be after the first passage.
