دورية أكاديمية
Purification and functional characterization of novel human skeletal stem cell lineages.
| العنوان: | Purification and functional characterization of novel human skeletal stem cell lineages. |
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| المؤلفون: | Hoover MY; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Ambrosi TH; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.; Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA.; Department of Orthopaedic Surgery, UC Davis Health, Sacramento, CA, USA., Steininger HM; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Koepke LS; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Wang Y; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.; Department of Orthopaedic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China., Zhao L; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.; Department of Orthopaedic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China., Murphy MP; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.; Blond McIndoe Laboratories, Division of Cell Matrix Biology and Regenerative Medicine, School of Biological Sciences, Faculty of Biology, Medicine and Health, The University of Manchester, Manchester Academic Health Science Centre, Manchester, UK., Alam AA; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Arouge EJ; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Butler MGK; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Takematsu E; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Stavitsky SP; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Hu S; Department of Orthopaedic Surgery, Stanford Hospitals and Clinics, Stanford, CA, USA., Sahoo D; Department of Pathology, University of California San Diego, La Jolla, CA, USA., Sinha R; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA., Morri M; Chan Zuckerberg BioHub, San Francisco, CA, USA.; Altos Labs, Redwood City, CA, USA., Neff N; Chan Zuckerberg BioHub, San Francisco, CA, USA., Bishop J; Department of Orthopaedic Surgery, Stanford Hospitals and Clinics, Stanford, CA, USA., Gardner M; Department of Orthopaedic Surgery, Stanford Hospitals and Clinics, Stanford, CA, USA., Goodman S; Department of Orthopaedic Surgery, Stanford Hospitals and Clinics, Stanford, CA, USA., Longaker M; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.; Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA.; Hagey Laboratory for Pediatric Regenerative Medicine, Stanford University School of Medicine, Stanford University, Stanford, CA, USA., Chan CKF; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA. chazchan@stanford.edu.; Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA. chazchan@stanford.edu.; Hagey Laboratory for Pediatric Regenerative Medicine, Stanford University School of Medicine, Stanford University, Stanford, CA, USA. chazchan@stanford.edu. |
| المصدر: | Nature protocols [Nat Protoc] 2023 Jul; Vol. 18 (7), pp. 2256-2282. Date of Electronic Publication: 2023 Jun 14. |
| نوع المنشور: | Journal Article; Review; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101284307 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1750-2799 (Electronic) Linking ISSN: 17502799 NLM ISO Abbreviation: Nat Protoc Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: London, UK : Nature Pub. Group, 2006- |
| مواضيع طبية MeSH: | Mesenchymal Stem Cells*, Humans ; Mice ; Animals ; Cell Lineage ; Reproducibility of Results ; Cell Differentiation/physiology ; Bone and Bones ; Bone Marrow Cells ; Cells, Cultured |
| مستخلص: | Human skeletal stem cells (hSSCs) hold tremendous therapeutic potential for developing new clinical strategies to effectively combat congenital and age-related musculoskeletal disorders. Unfortunately, refined methodologies for the proper isolation of bona fide hSSCs and the development of functional assays that accurately recapitulate their physiology within the skeleton have been lacking. Bone marrow-derived mesenchymal stromal cells (BMSCs), commonly used to describe the source of precursors for osteoblasts, chondrocytes, adipocytes and stroma, have held great promise as the basis of various approaches for cell therapy. However, the reproducibility and clinical efficacy of these attempts have been obscured by the heterogeneous nature of BMSCs due to their isolation by plastic adherence techniques. To address these limitations, our group has refined the purity of individual progenitor populations that are encompassed by BMSCs by identifying defined populations of bona fide hSSCs and their downstream progenitors that strictly give rise to skeletally restricted cell lineages. Here, we describe an advanced flow cytometric approach that utilizes an extensive panel of eight cell surface markers to define hSSCs; bone, cartilage and stromal progenitors; and more differentiated unipotent subtypes, including an osteogenic subset and three chondroprogenitors. We provide detailed instructions for the FACS-based isolation of hSSCs from various tissue sources, in vitro and in vivo skeletogenic functional assays, human xenograft mouse models and single-cell RNA sequencing analysis. This application of hSSC isolation can be performed by any researcher with basic skills in biology and flow cytometry within 1-2 days. The downstream functional assays can be performed within a range of 1-2 months. (© 2023. Springer Nature Limited.) |
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| معلومات مُعتمدة: | S10 OD028493 United States OD NIH HHS; K99 AG049958 United States AG NIA NIH HHS; R01 DE026730 United States DE NIDCR NIH HHS; U01 HL099999 United States HL NHLBI NIH HHS; U01 HL099776 United States HL NHLBI NIH HHS; R00 AG049958 United States AG NIA NIH HHS; I01 BX003754 United States BX BLRD VA; R01 DK115600 United States DK NIDDK NIH HHS; R01 CA086065 United States CA NCI NIH HHS; R01 HL058770 United States HL NHLBI NIH HHS; R21 DE019274 United States DE NIDCR NIH HHS; R01 AR063717 United States AR NIAMS NIH HHS; R56 DE025597 United States DE NIDCR NIH HHS; R01 DE021683 United States DE NIDCR NIH HHS; K99 AG066963 United States AG NIA NIH HHS; R21 DE024230 United States DE NIDCR NIH HHS; R01 DE027323 United States DE NIDCR NIH HHS; K08 GM069677 United States GM NIGMS NIH HHS; P50 HG007735 United States HG NHGRI NIH HHS; R00 AG066963 United States AG NIA NIH HHS; R01 AR055650 United States AR NIAMS NIH HHS; U24 DE026914 United States DE NIDCR NIH HHS |
| تواريخ الأحداث: | Date Created: 20230614 Date Completed: 20230712 Latest Revision: 20231108 |
| رمز التحديث: | 20231108 |
| مُعرف محوري في PubMed: | PMC10495180 |
| DOI: | 10.1038/s41596-023-00836-5 |
| PMID: | 37316563 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 1750-2799 |
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| DOI: | 10.1038/s41596-023-00836-5 |