Three-dimensional culture using human plasma-medium gel with fragmin/protamine microparticles for proliferation of various human cells |
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Authors: | Satoko Kishimoto Masayuki Ishihara Makoto Takikawa Megumi Takikawa Yuki Sumi Shingo Nakamura Masanori Fujita Toshinori Sato Tomoharu Kiyosawa |
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Institution: | 1. Research Support Center, Dokkyo Medical University, 880 Kitakobayashi, Mibu, Shimotsuga, 321-0293, Tochigi, Japan 2. Research Institute, National Defense Medical College, 3-2 Namiki, Tokorozawa, 359-8513, Saitama, Japan 3. Department of Medical Engineering, National Defense Medical College, Tokorozawa, Saitama, Japan 4. Department of Plastic Surgery, National Defense Medical College, Tokorozawa, Saitama, Japan 5. Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, Japan 6. Department of Biosciences and Informatics School of Fundamental Science and Technology, Keio University, Kanagawa, Japan
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Abstract: | Fragmin/protamine microparticles (F/P MPs) have been used as carriers for the preservation and activation of cytokines in human plasma (HP)–Dulbecco’s modified Eagle’s medium (DMEM) gels. This study investigated a three-dimensional (3D) culture system using an HP–DMEM gel with 0.1 mg/mL F/P MPs and 5 ng/mL FGF-2 for the proliferation of human dermal fibroblast cells (DFCs), human microvascular endothelial cells (MVECs) and human coronary smooth muscle cells (SMCs), or 5 ng/mL interleukin (IL)-3/granulocyte-macrophage colony-stimulating factor (GM-CSF) for a human hematopoietic cell line (TF-1 cells). DFCs, MVECs, SMCs and TF-1 cells grew rapidly under 3D culture conditions using a low-concentration HP (2 %)–DMEM gel with F/P MPs and FGF-2 (for DFCs, MVECs and SMCs) or IL-3/GM-CSF (for TF-1 cells) at doubling times of 22, 23, 25 and 18 h, respectively, without the use of animal serum, compared to under 2D culture conditions using low-concentration human serum (2 %)–DMEM with 5 ng/mL FGF-2 or IL-3/GM-CSF on F/P MP-coated plates at doubling times of approximately 26, 25, 40 and 20 h, respectively. |
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Keywords: | Cytokine carrier Fragmin/protamine microparticles (F/P MPs) Plasma–medium gel 3D culture |
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