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Enzyme‐Cleaved Bone Marrow Transplantation Improves the Engraftment of Bone Marrow Mesenchymal Stem Cells
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Enzyme‐Cleaved Bone Marrow Transplantation Improves the Engraftment of Bone Marrow Mesenchymal Stem Cells
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Journal Article
Enzyme‐Cleaved Bone Marrow Transplantation Improves the Engraftment of Bone Marrow Mesenchymal Stem Cells
2023
Overview
Mesenchymal stem cell (MSC) therapy is a promising approach to curing bone diseases and disorders. In treating genetic bone disorders, MSC therapy is local or systemic transplantation of isolated and in vitro proliferated MSC rather than bone marrow transplantation. Recent evidence showed that bone marrow MSC engraftment to bone regeneration has been controversial in animal and human studies. Here, our modified bone marrow transplantation (BMT) method solved this problem. Like routine BMT, our modified method involves three steps: (i) isolation of bone marrow cells from the donor, (ii) whole‐body lethal irradiation to the recipient, and (iii) injection of isolated bone marrow cells into irradiated recipient mice via the tail vein. The significant modification is imported at the bone marrow isolation step. While the bone marrow cells are flushed out from the bone marrow with the medium in routine BMT, we applied the enzymes’ (collagenase type 4 and dispase) integrated medium to wash out the bone marrow cells. Then, cells were incubated in enzyme integrated solution at 37°C for 10 minutes. This modification designated BMT as collagenase‐integrated BMT (c‐BMT). Notably, successful engraftment of bone marrow MSC to the new bone formation, such as osteoblasts and chondrocytes, occurs in c‐BMT mice, whereas routine BMT mice do not recruit bone marrow MSC. Indeed, flow cytometry data showed that c‐BMT includes a higher proportion of LepR+, CD51+, or RUNX2+ non‐hematopoietic cells than BMT. These findings suggested that c‐BMT is a time‐efficient and more reliable technique that ensures the disaggregation and collection of bone marrow stem cells and engraftment of bone marrow MSC to the recipient. Hence, we proposed that c‐BMT might be a promising approach to curing genetic bone disorders. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. Enzyme‐cleaved bone marrow isolation favors the isolation of bone marrow mesenchymal stem cells along with hematopoietic stem cells. Isolation of bone marrow cells using a normal culture medium (upper panel) is difficult to separate the bone marrow cell aggregates containing bone marrow mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs). The filtration limits the bone marrow cell aggregates from passing through. Applying enzymes such as collagenase and dispase (lower panel) dissociates the aggregation of MSCs and HSCs into single cells and facilitates the filtration step.
Publisher
John Wiley & Sons, Inc,Oxford University Press
