High-efficiency immunomagnetic isolation of solid tissue-originated integrin-expressing adult stem cells
| Autor: | Aaron Palmon, Yoav Neumann, Ran David, Guy Krief, Doron J. Aframian, Raluca Stiubea-Cohen |
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| Rok vydání: | 2012 |
| Předmět: |
Male
Cell Survival medicine.drug_class Cell Biology Immunomagnetic separation Monoclonal antibody Sensitivity and Specificity Salivary Glands General Biochemistry Genetics and Molecular Biology Flow cytometry Rats Sprague-Dawley medicine Animals Humans Molecular Biology Integrin alpha6beta1 medicine.diagnostic_test Immunomagnetic Separation Cell Membrane Antibodies Monoclonal Affinity Labels Microbead (research) Cell sorting Flow Cytometry Molecular biology Rats Adult Stem Cells medicine.anatomical_structure Head and Neck Neoplasms Stem cell Adult stem cell |
| Zdroj: | Methods. 56:305-309 |
| ISSN: | 1046-2023 |
| DOI: | 10.1016/j.ymeth.2011.10.002 |
| Popis: | Isolation of highly pure specific cell types is crucial for successful adult stem cell-based therapy. As the number of such cells in adult tissue is low, an extremely efficient method is needed for their isolation. Here, we describe cell-separation methodologies based on magnetic-affinity cell sorting (MACS) MicroBeads with monoclonal antibodies against specific membrane proteins conjugated to superparamagnetic particles. Cells labeled with MACS MicroBeads are retained in a magnetic field within a MACS column placed in a MACS separator, allowing fast and efficient separation. Both positively labeled and non-labeled fractions can be used directly for downstream applications as the separated cell fractions remain viable with no functional impairment. As immunomagnetic separation depends on the interaction between a cell's membrane and the magnetically labeled antibody, separation of specific cells originating from solid tissues is more complex and demands a cell-dissociating pretreatment. In this paper, we detail the use of immunomagnetic separation for the purpose of regenerating damaged salivary gland (SG) function in animal and human models of irradiated head and neck cancer. Each year 500,000 new cases of head and neck cancer occur worldwide. Most of these patients lose SG function following irradiation therapy. SGs contain integrin α6β1-expressing epithelial stem cells. We hypothesized that these cells can be isolated, multiplied in culture and auto-implanted into the irradiated SGs to regenerate damaged SG function. |
| Databáze: | OpenAIRE |
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