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使用含 Fragmin/鱼精蛋白微颗粒的人血浆介质凝胶进行三维培养,以促进各种人类细胞的增殖。

Three-dimensional culture using human plasma-medium gel with fragmin/protamine microparticles for proliferation of various human cells.

机构信息

Research Support Center, Dokkyo Medical University, 880 Kitakobayashi, Mibu, Shimotsuga, 321-0293, Tochigi, Japan,

出版信息

Cytotechnology. 2014 Oct;66(5):791-802. doi: 10.1007/s10616-013-9628-5. Epub 2013 Aug 17.

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.

摘要

Fragmin/鱼精蛋白微颗粒(F/P MPs)已被用作在人血浆(HP)-杜尔贝科改良伊格尔培养基(DMEM)凝胶中保存和激活细胞因子的载体。本研究采用含有 0.1mg/mL F/P MPs 和 5ng/mL FGF-2 的 HP-DMEM 凝胶,构建了一个三维(3D)培养系统,用于人真皮成纤维细胞(DFCs)、人微血管内皮细胞(MVECs)和人冠状动脉平滑肌细胞(SMCs)的增殖,或用于人造血细胞系(TF-1 细胞)的 5ng/mL 白细胞介素(IL)-3/粒细胞-巨噬细胞集落刺激因子(GM-CSF)。在 3D 培养条件下,DFCs、MVECs、SMCs 和 TF-1 细胞在低浓度 HP(2%)-DMEM 凝胶中快速生长,其中含有 F/P MPs 和 FGF-2(用于 DFCs、MVECs 和 SMCs)或 IL-3/GM-CSF(用于 TF-1 细胞),倍增时间分别为 22、23、25 和 18h,而无需使用动物血清,与在含有 5ng/mL FGF-2 或 IL-3/GM-CSF 的低浓度人血清(2%)-DMEM 上用 F/P MP 包被的平板在 2D 培养条件下相比,倍增时间分别约为 26、25、40 和 20h。

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