High-Throughput Lipidomic and Transcriptomic Analysis to Compare SP2/0, CHO, and HEK-293 Mammalian Cell Lines

Yue Zhang, Deniz Baycin-Hizal, Amit Kumar, Joseph Priola, Michelle Bahri, Kelley M. Heffner, Miao Wang, Xianlin Han, Michael A. Bowen, Michael J. Betenbaugh

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

A combined lipidomics and transcriptomics analysis was performed on mouse myeloma SP2/0, Chinese hamster ovary (CHO), and human embryonic kidney (HEK) cells in order to compare widely used mammalian expression systems. Initial thin layer chromatography (TLC) analysis indicated that phosphatidylethanolamine (PE) and phosphatidylcholine (PC) were the major lipid components in all cell lines with lower amounts of sphingomyelin (SM) in SP2/0 compared to CHO and HEK, which was subsequently confirmed and expanded upon following mass spectrometry (MS) analysis. HEK contained 4-10-fold higher amounts of lyso phosphatidylethanolamine (LPE) and 2-4-fold higher amounts of lyso phosphatidylcholine (LPC) compared to SP2/0 and CHO cell lines. C18:1 followed by C16:1 were the main contributors to the difference in both LPE and LPC levels. Alternatively, the SP2/0 cell line exhibited 30-65-fold lower amounts of SM principally in the amount of 16:0. By mapping the transcriptomics data to KEGG pathways, we found expression levels of secretory phospholipase A2 (sPLA2), lysophospholipid acyltransferase (LPEAT), lysophosphatidylcholine acyltransferase (LPCAT), and lysophospholipase (LYPLA) can contribute to the differences in LPE and LPC. Sphingomyelin synthases (SMS) and sphingomyelin phosphodiesterase (SMase) enzymes may play roles in SM differences across the three cell lines. The results of this study provide insights that will aid the understanding of the physiological and secretory differences across recombinant protein production systems.

Original languageEnglish (US)
Pages (from-to)1477-1485
Number of pages9
JournalAnalytical Chemistry
Volume89
Issue number3
DOIs
StatePublished - Feb 7 2017
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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