Walsh E J, Moran A P
Department of Microbiology, University College, Galway, Ireland.
J Appl Microbiol. 1997 Jul;83(1):67-75. doi: 10.1046/j.1365-2672.1997.00164.x.
An evaluation of the ability of various solid and liquid media to support both growth and antigen expression, particularly lipopolysaccharide (LPS) expression, by Helicobacter pylori culture collection strains and clinical isolates was performed. Liquid-based basal media (brain heart infusion, Brucella broth, Mueller-Hinton broth and tryptone soya broth) supported the growth of strains, whereas solid basal media of the same formulation did not support growth. Optimal growth of all strains was obtained on solid and in liquid media containing blood. Supplemented solid media containing supplements other than blood supported growth but only to a small extent. In liquid media excluding blood, serum supplements enhanced growth and horse serum was found to be superior to fetal calf serum. In general, beta-cyclodextrin did not increase growth. Mueller-Hinton broth or tryptone soya broth containing horse serum and a nitrogen source such as yeast extract or proteose peptone no. 3 were found to give optimal growth of H. pylori in a blood-free environment. Strains after cultivation in liquid media, irrespective of composition, maintained production of high-molecular weight (mol. wt) LPS with an O side chain independent of medium composition, whereas subculturing on solid media resulted in production of low-mol. wt LPS. Expression of proteins differed in liquid and on solid media, particularly proteins of 57 and 60 kDa, but qualitatively no differences were observed upon supplementation of basal media.
对多种固体和液体培养基支持幽门螺杆菌培养收集菌株和临床分离株生长及抗原表达(特别是脂多糖(LPS)表达)的能力进行了评估。液体基础培养基(脑心浸液、布鲁氏菌肉汤、穆勒-欣顿肉汤和胰蛋白胨大豆肉汤)支持菌株生长,而相同配方的固体基础培养基不支持生长。所有菌株在含血的固体和液体培养基上均能实现最佳生长。含除血液以外其他添加剂的固体培养基能支持生长,但程度有限。在不含血液的液体培养基中,血清添加剂可促进生长,且发现马血清优于胎牛血清。总体而言,β-环糊精不会促进生长。发现含有马血清和氮源(如酵母提取物或3号蛋白胨)的穆勒-欣顿肉汤或胰蛋白胨大豆肉汤能在无血环境中使幽门螺杆菌实现最佳生长。在液体培养基中培养后的菌株,无论其组成如何,均能维持高分子量(mol. wt)且带有O侧链的LPS的产生,其产生与培养基组成无关,而在固体培养基上继代培养则导致低分子量LPS的产生。液体和固体培养基中蛋白质的表达有所不同,特别是57 kDa和60 kDa的蛋白质,但在基础培养基中添加添加剂后,在定性方面未观察到差异。