title: Three-dimensional culture of mouse bone marrow cells on stroma formed within a porous scaffold: influence of scaffold shape and cryopreservation of the stromal layer on expansion of haematopoietic progenitor cells.
abstract: This study's primary goal was to develop an effective ex vivo expansion method for haematopoietic cells. 3D culture of mouse bone marrow cells was performed in porous scaffolds using a sheet or cube shape. Bone marrow cells were cultured on bone marrow-derived stromal layers formed within the scaffolds and the effect of scaffold shape on the expansion of haematopoietic cells was examined. In some experiments, stromal layers within cubic scaffolds were frozen and then used to culture bone marrow cells after thawing. Results show that after comparison, total cell density and expansion of haematopoietic cells were greater in cultures using the cubic scaffold, suggesting that it was superior to the sheet-like scaffold for expanding haematopoietic cells. When cryopreserved stroma was used, it effectively supported the expansion of haematopoietic cells, and a greater expansion of haematopoietic cells [(erythroid and haematopoietic progenitor cells (HPCs)] was achieved than in cultures with stromal cells that had not been cryopreserved. Expansion of cells using cryopreserved stroma had several other advantages such as a shorter culture period than the conventional method, a stable supply of stromal cells, and ease of handling and scaling up. As a result, this is an attractive method for ex vivo expansion of haematopoietic stem cells (HSCs) and HPCs for clinical use.
authors.affiliation: Department of Biomedical Engineering, University of Tsukuba, Ibaraki, Japan. hmiyoshi@md.tsukuba.ac.jp
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Miyoshi
authors.forename: Norio
authors.initials: N
authors.lastname: Ohshima
authors.forename: Chiaki
authors.initials: C
authors.lastname: Sato
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation; D002478:Cells, Cultured; D004396:Coloring Agents / Q000494:pharmacology; D015925:Cryopreservation / Q000379:methods; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning / Q000379:methods; D016062:Porosity; D013234:Stem Cells / Q000166:cytology; D017154:Stromal Cells / Q000166:cytology*; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue