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首页> 外文期刊>BioResearch open access. >Three-Dimensional Expansion Using Plasma-Medium Gel with Fragmin/Protamine Nanoparticles and FGF-2 to Stimulate Adipose-Derived Stromal Cells and Bone Marrow-Derived Mesenchymal Stem Cells
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Three-Dimensional Expansion Using Plasma-Medium Gel with Fragmin/Protamine Nanoparticles and FGF-2 to Stimulate Adipose-Derived Stromal Cells and Bone Marrow-Derived Mesenchymal Stem Cells

机译：使用等离子凝胶与Fragmin /鱼精蛋白纳米颗粒和FGF-2进行三维扩展，以刺激脂肪来源的基质细胞和骨髓来源的间充质干细胞

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Fragmin/protamine nanoparticles (F/P NPs) have been used as carriers for the preservation and controlled release of fibroblast growth factor (FGF)-2 and various cytokines in human plasma (HP). This study tested an HP–Dulbecco's modified Eagle's medium (DMEM) gel as a three-dimensional (3D) culture for the expansion of adipose tissue-derived multilineage stromal cells (ASCs) and bone marrow-derived mesenchymal stem cells (BMSCs). The growth of these cells improved in 3D culture using low-concentration HP (2%)–DMEM gel with 0.1?mg/mL F/P NPs and 5?ng/mL FGF-2 without animal serum in comparison to two-dimensional (2D) culture using a low-concentration human serum (2%)–DMEM containing 5?ng/mL FGF-2 on F/P NPs-coated plates. ASCs and BMSCs, which were expanded in the low-concentration HP–DMEM gel with F/P NPs and FGF-2, maintained their multilineage potential for differentiation into adipocytes or osteoblasts similar to the 2D cultured cells. Furthermore, flow cytometric analyses showed that the phenotypic markers which were positive for CD44, CD90, and CD105 (80%) and negative for CD34 and CD45 (1%) were well maintained in both 2D and 3D cultures after 7 days. Thus, this 3D culture system in low-concentration HP–DMEM gel with F/P NPs and FGF-2 provided an effective and safe method for the expansion of both cell types without using animal serum.
机译：Fragmin /鱼精蛋白纳米颗粒（F / P NPs）已被用作载体，用于在人血浆（HP）中保存和控制释放成纤维细胞生长因子（FGF）-2和各种细胞因子。这项研究测试了HP-Dulbecco改良的Eagle培养基（DMEM）凝胶作为三维（3D）培养物，以扩增脂肪组织衍生的多谱系基质细胞（ASC）和骨髓衍生的间充质干细胞（BMSC）。与二维分析相比，使用低浓度HP（2％）– DMEM凝胶，不含动物血清的0.1？mg / mL F / P NP和5？ng / mL FGF-2的3D培养，这些细胞的生长得以改善。 2D）在F / P NPs包被板上使用含有5？ng / mL FGF-2的低浓度人血清（2％）– DMEM进行培养。在具有F / P NP和FGF-2的低浓度HP-DMEM凝胶中扩增的ASC和BMSC，保持了其多谱系分化成类似于2D培养细胞的脂肪细胞或成骨细胞的潜力。此外，流式细胞术分析表明，在7天后，在2D和3D培养中，对CD44，CD90和CD105阳性（> 80％）而对CD34和CD45阴性（<1％）的表型标记均得到很好的维持。因此，这种具有F / P NP和FGF-2的低浓度HP-DMEM凝胶中的3D培养系统为不使用动物血清而扩增两种细胞提供了一种有效而安全的方法。

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