Recombinant collagen for animal product-free dextran microcarriers.
Recombinant collagen for animal product-free dextran microcarriers.
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DOI：
10.1007/s11626-008-9139-4
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发表时间：
2008-11
影响因子：
2.1
通讯作者：
Varani, James
中科院分区：
文献类型：
--
作者：
Dame, Michael K.;Varani, James
疫苗和其他生物制品制造商正面临来自监管机构越来越大的压力，要求他们开发完全不含动物成分的生产方法。为了满足这一需求，必须找到替代目前市场上的细胞培养基质，主要是胶原蛋白或明胶。在本文中，我们测试了一些可能的替代品，包括重组胶原蛋白，100 kda的重组明胶片段和从I型胶原蛋白的细胞结合区衍生的肽。在单层培养中，小的15个氨基酸肽不支持人成纤维细胞的附着。在单层培养中，100 kda的明胶片段支持细胞附着，但活性明显低于完整的猪明胶。重组I型胶原蛋白与天然胶原蛋白一样成功地促进细胞附着，两者都比猪明胶更有效。基于这些数据，用相同的附着蛋白——猪明胶、天然胶原蛋白或重组胶原蛋白——处理右旋糖酐微球。在单层培养中观察到相同的趋势。重组胶原（以及天然胶原）的浓度低至0.01 μg/cm2时支持细胞在葡聚糖微球上的附着。用低水平的聚乙烯亚胺（阳离子部分）处理右旋糖酐，当与低浓度的重组胶原蛋白结合使用时，进一步增强了附着力。细胞附着增加的地方，增殖也随之增加。基于这些发现，我们有信心，一种完全重组的替代品可以取代目前微载体制剂中的明胶，而不会失去天然蛋白质提供的细胞生长益处。
Manufacturers of vaccines and other biologicals are under increasing pressure from regulatory agencies to develop production methods that are completely animal-component-free. In order to comply with this demand, alternative cell culture substrates to those now on the market, primarily collagen or gelatin, must be found. In this paper, we have tested a number of possible substitutes including recombinant collagen, a 100-kDa recombinant gelatin fragment and a peptide derived from a cell-binding region of type I collagen. The small 15-amino acid peptide did not support attachment of human fibroblasts in monolayer culture. The 100-kDa gelatin fragment supported cell attachment in monolayer culture, but was significantly less active than intact porcine gelatin. Recombinant type I collagen was as successful in promoting cell attachment as native collagen, and both were more effective than porcine gelatin. Based on these data, dextran microspheres were treated with the same attachment proteins—porcine gelatin, native collagen, or recombinant collagen. The same trends were observed as in monolayer culture. Concentrations of the recombinant collagen (as well as native collagen) supported cell attachment on dextran microspheres at concentrations as low as 0.01 μg/cm2. Treatment of the dextran with a low level of polyethylenimine, a cationic moiety, further enhanced attachment when used in conjunction with the low concentration of recombinant collagen. Where there was increased cell attachment, increased proliferation followed. We are confident, based on these findings, that a fully recombinant substitute could replace gelatin in current microcarrier preparations without losing the cell growth benefits provided by the native protein.
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影响因子：
2.9
作者：
Ahlemeyer, B;Fischer, D;Krieglstein, J
通讯作者：
Krieglstein, J
影响因子：
2.2
作者：
Varani, J;Piel, F;Hillegas, WJ
通讯作者：
Hillegas, WJ
影响因子：
3.7
作者：
Fischer, D;Bieber, T;Kissel, T
通讯作者：
Kissel, T
影响因子：
4.1
作者：
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通讯作者：
Barabino, Gilda A.
影响因子：
4.9
作者：
Evans, MDM;Pavon-Djavid, G;Migonney, W
通讯作者：
Migonney, W