Functional and morphological characteristics of bovine adrenal chromaffin cells on macroporous poly(D,L-lactide-co-glycolide) scaffolds.
Functional and morphological characteristics of bovine adrenal chromaffin cells on macroporous poly(D,L-lactide-co-glycolide) scaffolds.
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大孔聚（D，L-丙交酯-乙交酯）支架上牛肾上腺嗜铬细胞的功能和形态特征。
DOI：
10.1089/107632703322495682
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Pappas,GeorgeD
中科院分区：
文献类型：
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作者：
Elcin,YMurat;Elcin,AEser;Pappas,GeorgeD
肾上腺染色质细胞（ACCs）分泌几种神经活性物质，这些物质有效地影响中枢神经系统的疼痛敏感性，并促进帕金森病患者内在黑质纹状体多巴胺能系统的恢复。ACC移植可能会因使用三维支架而上调。在这项研究中，我们确定了可生物降解的聚（D， l -乳酸-羟基乙酸）（PLGA）（85:15）海绵是否可以作为嗜铬细胞的载体。ACCs通过标准灌注（95%纯度）从牛肾上腺中分离出来，然后进行额外纯化（>纯度为99.5%）。将ACC （~5 × 105个细胞）混悬液（ⅰ型胶原）接种于预湿海绵上，在DMEM-F12（1:1）培养基（5%胎牛血清）中培养。采用高效液相色谱法和放射免疫法测定样品中儿茶酚胺和脑啡肽的含量。透射电镜观察细胞形态。形态学证据表明，在孔径为250 ~ 400 μm的支架上，染色质细胞的存活时间较长。细胞计数和扫描电镜显示，大部分种子细胞位于支架内。与单层培养相比，PLGA支架上的染色质细胞表现出更高水平的脑啡肽和儿茶酚胺。通过使用3d PLGA作为ACCs的支持，可以上调代谢功能并定位大量形态上看起来健康的细胞。在PLGA支架上培养的高纯度acc可能在移植研究中有希望，因为这些细胞的免疫原性较低，可以通过短期免疫抑制应用于体内环境。
Adrenal chromaffin cells (ACCs) secrete several neuroactive substances that are effective in influencing pain sensitivity in the central nervous system as well as enhancing the recovery of the intrinsic nigrostriatal dopaminergic system in patients with Parkinson's disease. ACC transplantation may be upregulated by the use of three-dimensional (3-D) scaffolds. In this study, we determined whether biodegradable poly(D,L-lactic-coglycolic acid) (PLGA) (85:15) sponges could be used as support for chromaffin cells. ACCs were isolated from bovine adrenal glands by standard perfusion (95% purity) followed by additional purification (>99.5% purity). ACC (~5 × 105cells) suspension in collagen (type I) was seeded on prewetted sponges and cultured in DMEM-F12 (1:1) medium (5% fetal bovine serum). The catecholamine and enkephalin levels of the samples were measured by high-performance liquid chromatography and radioimmunoassay. Cell morphology was examined by transmission electron microscopy. Morphological evidence showed prolonged viability of chromaffin cells on scaffolds having pores of 250-400 μm. Cell counts and scanning electron microscopy demonstrated that the majority of seeded cells were located within the scaffold. Chromaffin cells exhibited higher levels of enkephalins and catecholamines on PLGA scaffold compared with their monolayer cultures. By the use of 3-D PLGA as support for ACCs, it is possible to upregulate metabolic function and localize a high number of morphologically healthy-looking cells. Highly purified ACCs cultured on PLGA scaffold may have promise in transplantation studies, because these cells are less immunogenic and may be applied toin vivosettings by using short-term immunosuppression.