Isolation, culture and evaluation of multilineage-differentiating stress-enduring (Muse) cells

Yasumasa Kuroda, Shohei Wakao, Masaaki Kitada, Toru Murakami, Makoto Nojima, Mari Dezawa

Research output: Contribution to journalArticlepeer-review

138 Citations (Scopus)


Multilineage-differentiating stress-enduring (Muse) cells are distinct stem cells in mesenchymal cell populations with the capacity to self-renew, to differentiate into cells representative of all three germ layers from a single cell, and to repair damaged tissues by spontaneous differentiation into tissue-specific cells without forming teratomas. We describe step-by-step procedures for isolating and evaluating these cells. Muse cells are also a practical cell source for human induced pluripotent stem (iPS) cells with markedly high generation efficiency. They can be collected as cells that are double positive for stage-specific embryonic antigen-3 (SSEA-3) and CD105 from commercially available mesenchymal cells, such as adult human bone marrow stromal cells and dermal fibroblasts, or from fresh adult human bone marrow samples. Under both spontaneous and induced differentiation conditions, they show triploblastic differentiation. It takes 4-6 h to collect and 2 weeks to confirm the differentiation and self-renewal capacity of Muse cells.

Original languageEnglish
Pages (from-to)1391-1415
Number of pages25
JournalNature Protocols
Issue number7
Publication statusPublished - 2013 Jul


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