Comprehensive analysis of the secretome and transcriptome stability from WJ-Mesenchymal Stromal cells throughout GMP-Manufacturing

Mesenchymal stromal cells (MSCs) hold promise for cell-based therapies due to their ability to stimulate tissue repair and modulate immune responses. Umbilical cord-derived MSC from Wharton’s jelly (WJ), offer advantages such as low immunogenicity and potent immune modulatory effects. However, ensuring consistent quality and safety throughout their manufacturing process remains critical. RNA sequencing (RNA-seq) emerges as a crucial tool for assessing genetic stability and expression dynamics. This study examines the secretome and transcriptome signatures throughout WJ-MSCs Good Manufacturing Practice (GMP) production, focusing in the role of Total RNA or Massive Analysis of cDNA Ends (MACE-Seq)-RNA sequencing. Through comprehensive transcriptomic analysis, we demonstrated the stability of WJ-MSC transcriptome across manufacturing stages. Notably, MACE-Seq enhanced the identification of key expression patterns related to senescence and immunomodulation. These findings highlight the potential of MACE-Seq as a routine quality assessment tool for WJ-MSC-based therapies, ensuring their efficacy and safety in clinical applications. Importantly, MACE-Seq proves valuable for characterizing WJ-MSC products, providing insights unattainable through traditional assays.