Generation and differentiation of microtissues from multipotent precursor cells for use in tissue engineering
| Autor: | Michael Hentschel, Karin Berr, Jörg Handschel, Norbert R. Kübler, Christian Naujoks, Andrea Hassel, Gesine Kögler, Hans-Peter Wiesmann, Rita Depprich, Ulrich Meyer, Fabian Langenbach |
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| Rok vydání: | 2011 |
| Předmět: |
Staining and Labeling
Tissue Engineering Multipotent Stem Cells Cell Cell Culture Techniques Biology Fetal Blood Molecular biology General Biochemistry Genetics and Molecular Biology In vitro Microspheres medicine.anatomical_structure Tissue engineering Multipotent Stem Cell Cell culture Osteogenesis Precursor cell medicine Humans RNA RNA extraction Adult stem cell Biomedical engineering |
| Zdroj: | Nature protocols. 6(11) |
| ISSN: | 1750-2799 |
| Popis: | This protocol describes an effective method for the production of spherical microtissues (microspheres), which can be used for a variety of tissue-engineering purposes. The obtained microtissues are well suited for the study of osteogenesis in vitro when multipotent stem cells are used. The dimensions of the microspheres can easily be adjusted according to the cell numbers applied in an individual experiment. Thus, microspheres allow for the precise administration of defined cell numbers at well-defined sites. Here we describe a detailed workflow for the production of microspheres using unrestricted somatic stem cells from human umbilical cord blood and adapted protocols for the use of these microspheres in histological analysis. RNA extraction methods for mineralized microtissues are specifically modified for optimum yields. The duration of running the complete protocol without preparatory cell culture but including 2 weeks of microsphere incubation, histological staining and RNA isolation is about 3 weeks. |
| Databáze: | OpenAIRE |
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