Logo image
Back
Transient HA-100 exposure improves aggregate uniformity and cell-cell contact stability in suspension human pluripotent stem cell cultures
Journal article   Open access   Peer reviewed

Transient HA-100 exposure improves aggregate uniformity and cell-cell contact stability in suspension human pluripotent stem cell cultures

Preeti Khurana, Amar J. Azad, Nikola Kolundzic, Mirjana Liovic, Iustina F. Ivan, Jakob Jeriha, Norah M.E. Fogarty, Caroline Ogilvie, Anna Celli, Xiao Huang, …
Cytotherapy (Oxford, England), Forthcoming
Apr 2026
DOI: https://doi.org/10.1016/j.jcyt.2026.102904
Featured in Collection :   Drexel's Newest Publications
url
https://doi.org/10.1016/j.jcyt.2026.102904View
Published, Version of Record (VoR) Open

Abstract

aggregate uniformity HA-100 human pluripotent stem cells manufacturing suspension culture TJP1
Human pluripotent stem cell (hPSC) manufacturing workflows frequently rely on suspension aggregation, yet inter-line and batch-to-batch variability in aggregate formation can compromise process consistency and downstream differentiation performance. We evaluated whether a short exposure to HA-100, a small-molecule inhibitor of protein kinase A and protein kinase C signaling, could be used as an upstream process intervention to improve aggregate uniformity without compromising hPSC identity or developmental competence. Nine hPSC lines, including human embryonic stem cell and induced pluripotent stem cell lines, were examined in suspension culture. HA-100 treatment for the first 24 h promoted more compact and spherical aggregates, increased aggregate size into a narrower range across lines, and reduced overall variability relative to medium alone. Across the nine-line panel, HA-100-treated aggregates fell within an empirically defined size range of 25.37–33.95 × 10^-4 mm^3 after 24 h of suspension culture, providing a practical benchmark for process monitoring. To investigate the cellular basis of this effect, we generated an mCherry-TJP1 reporter hESC line, which enabled live visualization of junction dynamics. In calcium-depleted conditions, HA-100 delayed disruption of intercellular contacts and accelerated recovery of transepithelial electrical resistance, consistent with improved junctional resilience. Importantly, transient exposure to HA-100 did not abolish pluripotency marker expression or tri-lineage differentiation capacity. These data support HA-100 as a practical upstream intervention to reduce aggregate heterogeneity in suspension hPSC cultures and support manufacturing-oriented workflows that require greater reproducibility at the aggregation step.

Metrics

1 Record Views

Details

Logo image