In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islands
Abstract: The pursuit of ex vivo erythrocyte generation has led to the development of various culture systems that simulate the bone marrow microenvironment. However, these models often fail to fully replicate the hematopoietic niche's complex dynamics. In our research, we use a comprehensive s...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-05-01
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| Series: | Blood Advances |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2473952925001077 |
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| author | Christian A. Di Buduo Francesca Careddu Samuele Metti Marco Lunghi Santo Diprima Virginia Camilotto Giovanna Bruni Umberto Gianelli Delfina Tosi Cesare Perotti Claudia Del Fante Mario Cazzola Paola Braghetta David L. Kaplan Giampaolo Minetti Luca Malcovati Alessandra Balduini |
| author_facet | Christian A. Di Buduo Francesca Careddu Samuele Metti Marco Lunghi Santo Diprima Virginia Camilotto Giovanna Bruni Umberto Gianelli Delfina Tosi Cesare Perotti Claudia Del Fante Mario Cazzola Paola Braghetta David L. Kaplan Giampaolo Minetti Luca Malcovati Alessandra Balduini |
| author_sort | Christian A. Di Buduo |
| collection | DOAJ |
| description | Abstract: The pursuit of ex vivo erythrocyte generation has led to the development of various culture systems that simulate the bone marrow microenvironment. However, these models often fail to fully replicate the hematopoietic niche's complex dynamics. In our research, we use a comprehensive strategy that emphasizes physiological red blood cell (RBC) differentiation using a minimal cytokine regimen. A key innovation in our approach is the integration of a 3-dimensional (3D) silk-based scaffold engineered to mimic both the physical and chemical properties of human bone marrow. This scaffold facilitates critical macrophage-RBC interactions and incorporates fibronectin functionalization to support the formation of erythroblastic island (EBI)–like niches. We observed diverse stages of erythroblast maturation within these niches, driven by the activation of autophagy, which promotes organelle clearance and membrane remodeling. This process leads to reduced surface integrin expression and significantly enhances RBC enucleation. Using a specialized bioreactor chamber, millions of RBCs can be detached from the EBIs and collected in transfusion bags via dynamic perfusion. Inhibition of autophagy through pharmacological agents or α4 integrin blockade disrupted EBI formation, preventing cells from completing their final morphological transformations, having them trapped in the erythroblast stage. Our findings underscore the importance of the bone marrow niche in maintaining the structural integrity of EBIs and highlight the critical role of autophagy in facilitating organelle clearance during RBC maturation. RNA sequencing analysis further confirmed that these processes are uniquely supported by the 3D silk scaffold, which is essential for enhancing RBC production ex vivo. |
| format | Article |
| id | doaj-art-7560fdce919a4b05ab22145479dfdab7 |
| institution | OA Journals |
| issn | 2473-9529 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Blood Advances |
| spelling | doaj-art-7560fdce919a4b05ab22145479dfdab72025-08-20T02:19:47ZengElsevierBlood Advances2473-95292025-05-01992192220610.1182/bloodadvances.2024014905In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islandsChristian A. Di Buduo0Francesca Careddu1Samuele Metti2Marco Lunghi3Santo Diprima4Virginia Camilotto5Giovanna Bruni6Umberto Gianelli7Delfina Tosi8Cesare Perotti9Claudia Del Fante10Mario Cazzola11Paola Braghetta12David L. Kaplan13Giampaolo Minetti14Luca Malcovati15Alessandra Balduini16Department of Molecular Medicine, University of Pavia, Pavia, ItalyDepartment of Molecular Medicine, University of Pavia, Pavia, ItalyDepartment of Molecular Medicine, University of Padova, Padova, ItalyDepartment of Molecular Medicine, University of Pavia, Pavia, ItalyBioinformatic Division, Center for Omics Sciences, Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele Scientific Institute, Milan, ItalyDepartment of Molecular Medicine, University of Pavia, Pavia, Italy; Department of Hematology Oncology, Hematology Unit, Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo, Pavia, ItalyCenter for Colloid and Surface Science-Department of Chemistry, Physical-Chemistry Section, University of Pavia, Pavia, ItalyDepartment of Health Sciences, University of Milan, Struttura Complessa di Anatomia Patologica, Azienda Socio-Sanitaria Territoriale-Santi Paolo e Carlo, Milan, ItalyDepartment of Health Sciences, University of Milan, Struttura Complessa di Anatomia Patologica, Azienda Socio-Sanitaria Territoriale-Santi Paolo e Carlo, Milan, ItalyDivision of Immunohaematology and Transfusion Service, Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo, Pavia, ItalyDivision of Immunohaematology and Transfusion Service, Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo, Pavia, ItalyDepartment of Molecular Medicine, University of Pavia, Pavia, Italy; Department of Hematology Oncology, Hematology Unit, Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo, Pavia, ItalyDepartment of Molecular Medicine, University of Padova, Padova, ItalyDepartment of Biomedical Engineering, Tufts University, Medford, MADepartment of Biology and Biotechnology “Lazzaro Spallanzani,” University of Pavia, Pavia, ItalyDepartment of Molecular Medicine, University of Pavia, Pavia, Italy; Department of Hematology Oncology, Hematology Unit, Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo, Pavia, ItalyDepartment of Molecular Medicine, University of Pavia, Pavia, Italy; Department of Biomedical Engineering, Tufts University, Medford, MA; Correspondence: Alessandra Balduini, Department of Molecular Medicine, University of Pavia, Viale Golgi n. 19, 27100, Pavia, Italy;Abstract: The pursuit of ex vivo erythrocyte generation has led to the development of various culture systems that simulate the bone marrow microenvironment. However, these models often fail to fully replicate the hematopoietic niche's complex dynamics. In our research, we use a comprehensive strategy that emphasizes physiological red blood cell (RBC) differentiation using a minimal cytokine regimen. A key innovation in our approach is the integration of a 3-dimensional (3D) silk-based scaffold engineered to mimic both the physical and chemical properties of human bone marrow. This scaffold facilitates critical macrophage-RBC interactions and incorporates fibronectin functionalization to support the formation of erythroblastic island (EBI)–like niches. We observed diverse stages of erythroblast maturation within these niches, driven by the activation of autophagy, which promotes organelle clearance and membrane remodeling. This process leads to reduced surface integrin expression and significantly enhances RBC enucleation. Using a specialized bioreactor chamber, millions of RBCs can be detached from the EBIs and collected in transfusion bags via dynamic perfusion. Inhibition of autophagy through pharmacological agents or α4 integrin blockade disrupted EBI formation, preventing cells from completing their final morphological transformations, having them trapped in the erythroblast stage. Our findings underscore the importance of the bone marrow niche in maintaining the structural integrity of EBIs and highlight the critical role of autophagy in facilitating organelle clearance during RBC maturation. RNA sequencing analysis further confirmed that these processes are uniquely supported by the 3D silk scaffold, which is essential for enhancing RBC production ex vivo.http://www.sciencedirect.com/science/article/pii/S2473952925001077 |
| spellingShingle | Christian A. Di Buduo Francesca Careddu Samuele Metti Marco Lunghi Santo Diprima Virginia Camilotto Giovanna Bruni Umberto Gianelli Delfina Tosi Cesare Perotti Claudia Del Fante Mario Cazzola Paola Braghetta David L. Kaplan Giampaolo Minetti Luca Malcovati Alessandra Balduini In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islands Blood Advances |
| title | In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islands |
| title_full | In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islands |
| title_fullStr | In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islands |
| title_full_unstemmed | In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islands |
| title_short | In vitro studies of human erythropoiesis using a 3D silk-based bone marrow model that generates erythroblastic islands |
| title_sort | in vitro studies of human erythropoiesis using a 3d silk based bone marrow model that generates erythroblastic islands |
| url | http://www.sciencedirect.com/science/article/pii/S2473952925001077 |
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