title: Polyurethane scaffolds seeded with CD34(+) cells maintain early stem cells whilst also facilitating prolonged egress of haematopoietic progenitors.
abstract: We describe a 3D erythroid culture system that utilises a porous polyurethane (PU) scaffold to mimic the compartmentalisation found in the bone marrow. PU scaffolds seeded with peripheral blood CD34(+) cells exhibit a remarkable reproducibility of egress, with an increased output when directly compared to human bone scaffolds over 28 days. Immunofluorescence demonstrated the persistence of CD34(+) cells within the scaffolds for the entirety of the culture. To characterise scaffold outputs, we designed a flow cytometry panel that utilises surface marker expression observed in standard 2D erythroid and megakaryocyte cultures. This showed that the egress population is comprised of haematopoietic progenitor cells (CD36(+)GPA(-/low)). Control cultures conducted in parallel but in the absence of a scaffold were also generally maintained for the longevity of the culture albeit with a higher level of cell death. The harvested scaffold egress can also be expanded and differentiated to the reticulocyte stage. In summary, PU scaffolds can behave as a subtractive compartmentalised culture system retaining and allowing maintenance of the seeded "CD34(+) cell" population despite this population decreasing in amount as the culture progresses, whilst also facilitating egress of increasingly differentiated cells.
authors: School of Biochemistry, Biomedical Sciences Building, University of Bristol, Bristol, BS8 1TD, United Kingdom.|National Institute for Health Research Blood and Transplant Research Unit (NIHR BTRU) in Red Blood Cell Products, University of Bristol, Bristol, BS8 1TD, United Kingdom.
authors: Charlotte E
authors: CE
authors: Severn
authors: Biological Systems Engineering Laboratory, Department of Chemical Engineering, Imperial College London, London, SW7 2AZ, United Kingdom.
authors: Hugo
authors: H
authors: Macedo
authors: Tissue and Eye Services, NHS Blood and Transplant, Liverpool, L24 8RB, United Kingdom.
authors: Mark J
authors: MJ
authors: Eagle
authors: Tissue and Eye Services, NHS Blood and Transplant, Liverpool, L24 8RB, United Kingdom.
authors: Paul
authors: P
authors: Rooney
authors: Biological Systems Engineering Laboratory, Department of Chemical Engineering, Imperial College London, London, SW7 2AZ, United Kingdom.
authors: Athanasios
authors: A
authors: Mantalaris
authors: School of Biochemistry, Biomedical Sciences Building, University of Bristol, Bristol, BS8 1TD, United Kingdom.|National Institute for Health Research Blood and Transplant Research Unit (NIHR BTRU) in Red Blood Cell Products, University of Bristol, Bristol, BS8 1TD, United Kingdom.
authors: Ashley M
authors: AM
authors: Toye
mesh_terms: D018952:Antigens, CD34; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D011140:Polyurethanes / Q000737:chemistry*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D018952:Antigens, CD34; D011140:Polyurethanes