Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409, USA.
Center of Biotechnology and Genomics, Texas Tech University, Lubbock, TX 79409, USA.
Biomolecules. 2022 Oct 27;12(11):1575. doi: 10.3390/biom12111575.
Aiming to reduce the gap between in vitro and in vivo environment, a complex culture medium, Plasmax, was introduced recently, which includes nutrients and metabolites with concentrations normally found in human plasma. Herein, to study the influence of this medium on cellular behaviors, we utilized Plasmax to cultivate two cancer cell lines, including one breast cancer cell line, MDA-MB-231BR, and one brain cancer cell line, CRL-1620. Cancer cells were harvested and prepared for transcriptomics and proteomics analyses to assess the discrepancies caused by the different nutritional environments of Plasmax and two commercial media: DMEM, and EMEM. Total RNAs of cells were extracted using mammalian total RNA extract kits and analyzed by next-generation RNA sequencing; proteomics analyses were performed using LC-MS/MS. Gene oncology and pathway analysis were employed to study the affected functions. The cellular invasion and cell death were inhibited in MDA-MB-231BR cell line when cultured in Plasmax compared to DMEM and EMEM, whereas the invasion, migration and protein synthesis of CRL-1620 cell line were activated in Plasmax in relative to both commercial media. The expression changes of some proteins were more significant compared to their corresponding transcripts, indicating that Plasmax has more influence upon regulatory processes of proteins after translation. This work provides complementary information to the original study of Plasmax, aiming to facilitate the selection of appropriate media for in vitro cancer cell studies.
为了缩小体外和体内环境之间的差距，最近引入了一种复杂的培养基 Plasmax，其中包含通常在人血浆中发现的营养物质和代谢物。在这里，为了研究该培养基对细胞行为的影响，我们使用 Plasmax 培养了两种癌细胞系，包括一种乳腺癌细胞系 MDA-MB-231BR 和一种脑癌细胞系 CRL-1620。收获癌细胞并进行转录组学和蛋白质组学分析，以评估 Plasmax 和两种商业培养基 DMEM 和 EMEM 的不同营养环境引起的差异。使用哺乳动物总 RNA 提取试剂盒提取细胞中的总 RNA，并通过下一代 RNA 测序进行分析；使用 LC-MS/MS 进行蛋白质组学分析。通过肿瘤基因和途径分析研究受影响的功能。与 DMEM 和 EMEM 相比，MDA-MB-231BR 细胞系在 Plasmax 中培养时，细胞侵袭和细胞死亡受到抑制，而 CRL-1620 细胞系的侵袭、迁移和蛋白质合成在 Plasmax 中相对于两种商业培养基被激活。与相应的转录物相比，一些蛋白质的表达变化更为显著，这表明 Plasmax 在翻译后对蛋白质的调节过程有更大的影响。这项工作为 Plasmax 的原始研究提供了补充信息，旨在为体外癌细胞研究选择合适的培养基提供便利。
