Greiner, Johannes ; Grunwald, Lena-Marie; Müller, Janine; Sudhoff, Holger ; Widera, Darius ; Kaltschmidt, Christian; Kaltschmidt, Barbara: Culture bag systems for clinical applications of adult human neural crest-derived stem cells. In: Stem Cell Research & Therapy. Jg.5 H. 2. 2014
Inhalt
- Introduction
- Methods
- Isolation and three-dimensional-cell culture of adult human inferior turbinate stem cells
- Cultivation of ITSCs in Afc-FEP bags
- Transfection of cultivated ITSCs
- Proliferation assay and vitality measurement of ITSCs
- Reverse transcription PCR
- Flow cytometric measurement of DNA content
- Telomerase activity measurement
- Directed osteogenic differentiation assay
- Detection of alkaline phosphatase-activity
- Alizarin Red S and Von Kossa staining for assessment of osteogenic differentiation
- Adipogenic differentiation assay
- Directed neuronal differentiation
- Immunocytochemistry of differentiated ITSCs
- Vesicle recycling
- Results
- Afc-FEP cell culture bags are suitable for three-dimensional-cultivation of human ITSCs
- Bag culture does not impair viability and proliferative capability of ITSCs
- ITSCs maintain their expression profile and genetic stability during bag-culture
- Subculture of ITSCs in bags does not affect their telomerase activity
- ITSCs cultivated in bags show efficient osteogenic differentiation capability in a directed differentiation assay
- ITSCs give rise to adipogenic cell types after bag-culture
- Bag-cultured ITSCs maintain their ability to differentiate into neuronal cells
- Discussion
- Conclusions
- Abbreviations
- Competing interests
- Authors’ contributions
- Acknowledgements
- Author details
- References