Isolation of therapeutically functional mouse bone marrow mesenchymal stem cells within 3 h by an effective single‐step plastic‐adherent method

作者：Felix Shih‐Hsiang Hsiao, C. C. Cheng, S.‐Y. Peng, Hsin‐Yu Huang, Wei‐Shiung Lian, M.‐L. Jan, Ya-Ching Fang, Enhao Cheng, Kyung Ho Lee, Wei-Jhih Cheng, Shau‐Ping Lin, Shinn‐Chih Wu · 发表于：Cell Proliferation · 年份：2010 · DOI：10.1111/j.1365-2184.2010.00674.x · 被引用次数：33 · 研究领域：Mesenchymal stem cell research、Hematopoietic Stem Cell Transplantation、Bone and Joint Diseases

OBJECTIVES: Isolation of mouse mesenchymal stem cells (mMSCs), by the approach of plastic adherence, has been difficult due to persistent contamination by haematopoietic cells (HCs); we have observed that this contamination was due to engagement between HCs and mMSCs. The HCs can be lifted together with the mMSCs despite their insensitivity to trypsin digestion. Herein, we provide a single-step procedure to rapidly segregate mMSCs from HC contaminants using transient lower-density plastic adherence (tLDA). MATERIALS AND METHODS: The tLDA was performed by replating bone marrow adherent cells at lower density (1.25 x 10(4) cells/cm(2)) than usual, allowing for transient adherence of no more than 3 h, followed by trypsin digestion. tLDA-isolated cells were evaluated by immunophenotyping, multi-differentiation potentials, immunosuppressive properties, and therapeutic potential as demonstrated by symptoms of osteoporosis. RESULTS: The single-step tLDA method can effectively eliminate the persistent HC contaminants; tLDA-isolated cells were phenotypically equivalent to those reported as mMSCs. The isolated cells possessed classic tri-lineage differentiation potential into osteogenic, adipogenic and chondrogenic lineages and had immunosuppressive properties. After intravenous transplantation, they migrated into the allogeneic bone marrow and rescued hosts from osteoporosis symptoms, demonstrating their therapeutic potential. CONCLUSIONS: We have developed a simple and economical met...