論文 - 詳細
| RRC ID | 57995 |
|---|---|
| 著者 | Sekai M, Wang J, Hamazaki Y. |
| タイトル | Clonogenic Culture of Mouse Thymic Epithelial Cells. |
| ジャーナル | Methods Mol Biol |
| Abstract |
The thymus plays an essential role in the development and selection of T cells by providing a unique microenvironment that is mainly composed of thymic epithelial cells (TECs). We previously identified stem cells of medullary TECs (mTECs) that are crucial for central tolerance induction using a novel clonogenic culture system. We also found that medullary thymic epithelial stem cells (mTESCs) maintain life-long mTECs regeneration and central T cell self-tolerance in mouse models. The clonogenic efficiency of TECs in vitro is highly correlated to the TEC reconstitution activity in vivo. Here, we describe the clonogenic culture system to evaluate the self-renewing activity of TESCs. The colonies are derived from TESCs, are visualized and quantified by rhodamine-B staining on a feeder layer, and can be passaged in vitro. Thus, our system enables quantitative evaluation of TESC activity and is useful for dissecting the mechanisms that regulate TESC activity in physiological aging as well as in various clinical settings. |
| 巻・号 | 2048 |
| ページ | 143-153 |
| 公開日 | 2019-1-1 |
| DOI | 10.1007/978-1-4939-9728-2_15 |
| PMID | 31396938 |
| MeSH | Aging / physiology Animals Cell Differentiation / physiology Cell Line Coculture Techniques / instrumentation Coculture Techniques / methods Colony-Forming Units Assay / instrumentation Colony-Forming Units Assay / methods* DNA-Binding Proteins / genetics Epithelial Cells / physiology Flow Cytometry / instrumentation Flow Cytometry / methods Fluorescent Antibody Technique, Direct / instrumentation Fluorescent Antibody Technique, Direct / methods Fluorescent Dyes / chemistry Immunomagnetic Separation / instrumentation Immunomagnetic Separation / methods Mice Mice, Knockout Primary Cell Culture / instrumentation Primary Cell Culture / methods* Rhodamines / chemistry Self Tolerance / physiology Staining and Labeling / methods Stem Cells / physiology Thymus Gland / cytology* |
| 引用数 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | STO(RCB0536) |