Reliable isolation of human mesenchymal stromal cells from bone marrow biopsy specimens in patients after allogeneic hematopoietic cell transplantation |
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| Affiliation: | 1. Department of Internal Medicine I, University Hospital Carl Gustav Carus, Dresden, Germany;2. German Cancer Consortium (DKTK), Partner Site Dresden, and German Cancer Research Center (DKFZ), Heidelberg, Germany;3. University Cancer Centrum (UCC), Early Clinical Trial Unit (ECTU), University Hospital Carl Gustav Carus, Dresden, Germany;4. Max Planck Institute for the Science of Light & Max-Planck-Zentrum für Physik und Medizin, Erlangen, Germany;5. Biotechnology Center, Center for Molecular and Cellular Bioengineering TU Dresden Tatzberg 47-49, Dresden, Germany;6. Institute of Immunology, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany;7. National Center for Tumor Diseases (NCT), Partner Site Dresden, Dresden, Germany;8. Center for Regenerative Therapies (CRTD), Dresden, Germany;1. Division of Hematology and Medical Oncology, Mayo Clinic, Jacksonville, Florida, USA;2. Department of Laboratory Medicine & Pathology, Mayo Clinic, Jacksonville, Florida, USA;3. Division of Internal Medicine-Clinical Hematology, Al-Azhar University, Cairo, Egypt;1. State Key Laboratory of Experimental Hematology, National Clinical Research Center for Hematological Disorders, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China;2. Department of Anesthesiology, Beijing Hospital, National Center of Gerontology, Beijing, China;3. Department of Pathology, Harvard Medical School, Dana-Farber/Harvard Cancer Center, Boston, Massachusetts, USA;1. Alberta Health Services, Edmonton, Alberta, Canada;2. Alberta Health Services, Calgary, Alberta, Canada;3. University of Calgary, Calgary, Alberta, Canada |
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| Abstract: | Isolation of mesenchymal stromal cells (MSCs) from pretreated, hematologic patients is challenging. Especially after allogeneic hematopoietic cell transplantation (HCT), standard protocols using bone marrow aspirates fail to reliably recover sufficient cell numbers. Because MSCs are considered to contribute to processes that mainly affect the outcome after transplantation, such as an efficient lymphohematopoietic recovery, extent of graft-versus-host disease as well as the occurrence of leukemic relapse, it is of great clinical relevance to investigate MSC function in this context. Previous studies showed that MSCs can be isolated by collagenase digestion of large bone fragments of hematologically healthy patients undergoing hip replacement or knee surgeries. We have now further developed this procedure for the isolation of MSCs from hematologic patients after allogeneic HCT by using trephine biopsy specimens obtained during routine examinations. Comparison of aspirates and trephine biopsy specimens from patients after allogeneic HCT revealed a significantly higher frequency of clonogenic MSCs (colony-forming unit–fibroblast [CFU-F]) in trephine biopsy specimens (mean, 289.8 ± standard deviation 322.5 CFU-F colonies/1 × 106 total nucleated cells versus 4.2 ± 9.9; P < 0.0001). Subsequent expansion of functional MSCs isolated from trephine biopsy specimen was more robust and led to a significantly higher yield compared with control samples expanded from aspirates (median, 1.6 × 106; range, 0–2.3 × 107 P0 MSCs versus 5.4 × 104; range, 0–8.9 × 106; P < 0.0001). Using trephine biopsy specimens as MSC source facilitates the investigation of various clinical questions. |
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