Calcium oxalate crystal attachment to cultured kidney epithelial cell lines.
Calcium oxalate crystal attachment to cultured kidney epithelial cell lines.
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DOI：
10.1016/s0022-5347(01)62607-7
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发表时间：
1998-10
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影响因子：
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通讯作者：
M. Bigelow;J. Wiessner;J. Kleinman;N. Mandel
中科院分区：
文献类型：
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作者：
M. Bigelow;J. Wiessner;J. Kleinman;N. Mandel
目的培养的肾上皮细胞系已广泛应用于尿石症的研究，尤其是结石晶体与细胞膜相互作用的研究。有证据表明，当上皮细胞系被转化或连续传代以使其永生化时，它们会经历细胞生理学和形态学的变化。由于缺乏自发性结石病的动物模型，利用细胞培养进行结石研究常常是必要的。然而，这些细胞培养研究的解释可能会被原代细胞和连续细胞培养之间的细胞生理学的任何显著差异所模糊。材料与方法研究的细胞系为原代小鼠近曲小管细胞（pMPT）、原代内髓集合管细胞（pIMCD）、半连续内髓集合管细胞（cIMCD）、BSC-1细胞、COS-1细胞、LLC-PK 1细胞、MDCK细胞、NRK-52 E细胞和OK细胞。所有的细胞系培养在相同的条件下，COM附件的量进行了测量，使用放射性标记COM crystals.ResultsCOM晶体与连续肾上皮细胞的相互作用不同的两个因素之间的不同的细胞系。一般来说，生长为规则的、汇合的细胞单层的细胞，如pMPT、pIMCD和cIMCD细胞，表现出最低水平的晶体附着。无论是膜流动性的变化，也没有损失正常上皮细胞膜的不对称性似乎与晶体附着。连续细胞培养9天后，COM与cIMCD细胞的附着下降了61%，而与MDCK细胞的晶体附着保持不变。目前还不清楚是什么使这些细胞系更耐晶体attachment相比，连续celllines.ConclusionsThe显着差异COM附件之间的原代肾上皮细胞和连续上皮细胞培养物和生长形态之间的明显差异，必须考虑当选择一个细胞系用于肾结石的研究。比较cIMCD细胞和MDCK细胞在延长的培养时间揭示了COM附着的差异的一个可能的解释：成熟的，终末分化的，非分裂细胞单层的形成可以保护细胞免于晶体附着。
PurposeCultured kidney epithelial cell lines have frequently been used in urolithiasis research, and in particular in studies related to the interactions between stone crystals and cell membranes. There is evidence that when epithelial cell lines are transformed or serially passed to immortalize them, they experience changes in both cell physiology and morphology. Stone research utilizing cell cultures is frequently necessary due to the lack of an animal model for spontaneous stone disease. However, the interpretation of these cell culture research studies might be clouded by any significant differences in cell physiology between primary cells and continuous cell cultures. Therefore, the present study was conducted to compare calcium oxalate monohydrate (COM) crystal attachment to two primary kidney epithelial cell lines and to various continuous cell lines.Materials and MethodsThe cell lines surveyed were primary mouse proximal tubule cells (pMPT), primary inner medullary collecting duct cells (pIMCD), semi-continuous inner medullary collecting duct cells (cIMCD), BSC-1 cells, COS-1 cells, LLC-PK1cells, MDCK cells, NRK-52E cells, and OK cells. All cell lines were cultured under identical conditions and the amount of COM attachment was measured using radioactive labeled COM crystals.ResultsCOM crystal interaction with continuous kidney epithelial cells varied by a factor of two among the different cell lines. In general, cells that grew as regular, confluent cell monolayers, such as pMPT, pIMCD and cIMCD cells, exhibited the lowest levels of crystal attachment. Neither changes in membrane fluidity nor loss of normal epithelial cell membrane asymmetry seemed to correlate well with crystal attachment. After nine days of continuous cell culture, COM attachment to cIMCD cells dropped by 61 percent while crystal attachment to MDCK cells remained unchanged. It is unclear what makes these cell lines more resistant to crystal attachment compared to continuous cell lines.ConclusionsThe significant difference in COM attachment between primary kidney epithelial cells and continuous epithelial cell cultures and the apparent differences in growth morphology between primary and continuous cell cultures must be considered when selecting a cell line for use in kidney stone research. Comparison of cIMCD cells and MDCK cells during extended culture time revealed one possible explanation for the differences in COM attachment: the formation of a mature, end-differentiated, non-dividing cell monolayer could protect the cells from crystal attachment.