Evaluation of serum-free media formulations in feeder cell–stimulated expansion of natural killer cells

Jena E. Moseman, Jennifer A. Foltz, Kinnari Sorathia, Erica L. Heipertz, Dean A. Lee

Research output: Contribution to journalArticlepeer-review

14 Scopus citations


Background: Optimal expansion of therapeutic natural killer (NK) cell products has required media supplementation with human or fetal bovine serum, which raises safety and regulatory concerns for clinical manufacturing. Serum-free media (SFM) have been optimized for T-cell expansion, but few SFM systems have been developed for NK cells. Here, we compare six commercial clinical-grade SFM with our standard fetal bovine serum–containing medium for their ability to support NK cell expansion and function. Methods: Human peripheral blood NK cells were expanded in selected media by recursive weekly stimulation with K562-based feeder cells expressing membrane-bound interleukin-21 and CD137L. Expansion was the primary readout, and the best-performing SFM was then compared with standard medium for cytotoxicity, phenotype, degranulation and cytokine secretion. Multiple lots were compared for consistency, and media was analyzed throughout for nutrient consumption and metabolic byproducts. Results: TexMACS, OpTmizer, SCGM, ABS-001 and StemXVivo demonstrated equal or inferior NK cell expansion kinetics compared with standard medium, but expansion was markedly superior with AIM V + 5% Immune Cell Serum Replacement (ICSR; mean 5448 vs. 2621-fold expansion in 14 days). Surprisingly, NK cells expanded in AIM V + ICSR also showed increased cytotoxicity, tumor necrosis factor α secretion and DNAM-1, NKG2D, NKp30, FasL, granzyme B and perforin expression. Lot-to-lot variability was minimal. Glucose and glutamine consumption were inversely related to lactate and ammonia production. Discussion: The AIM V + ICSR SFM system supports excellent ex vivo expansion of clinical-grade NK cells with the phenotype and function needed for adoptive immunotherapy.

Original languageEnglish
Pages (from-to)322-328
Number of pages7
Issue number6
StatePublished - Jun 2020


  • cancer immunotherapy
  • natural killer cells
  • serum-free medium


Dive into the research topics of 'Evaluation of serum-free media formulations in feeder cell–stimulated expansion of natural killer cells'. Together they form a unique fingerprint.

Cite this