The human amniotic membrane derived MSC secretome restrains neuroinflammation and peripheral immune cell invasion of 3D neural tissue through conserved CXCL10 blockade and proteostatic rescue

Chronic neuroinflammation is a convergent mechanism of neurodegenerative disease progression, yet therapies that restrain it without impairing central nervous system immune surveillance remain limited. The human amniotic membrane-derived mesenchymal stromal cell secretome (CM-hAMSC) was shown to modulate peripheral immunity, with demonstrated efficacy in several inflammatory models, but its action on the neuroinflammatory circuitry is poorly defined. Here, we systematically investigated the effects of CM-hAMSC across a nested hierarchy of complementary neuroinflammatory models, including: monolayer cultures of human cell line-derived neurons, astrocytes and microglia; 3D human tri-partite neurospheroids; a peripheral immune transwell interface; and microglia-free murine induced pluripotent stem cell-derived bi-partite neurospheroids, the latter of which was profiled by unbiased transcriptomics and proteomics. CM-hAMSC suppressed pro-inflammatory and cell death programs while preserving neuronal transcriptional programs associated with physiological function. Selectively modulated peripheral immune recruitment through suppression of CD4⁺ T-cell migration while sparing CD8⁺ T cells, prevented stress granule assembly in neurons and astrocytes, and markedly reduced syngeneic splenocyte invasion of inflamed 3D neural tissue. Integrated multi-omics revealed the inhibition of death and stress pathways and the concomitant upregulation of pro-survival pathways. Across four independent experimental systems, suppression of CXCL10 emerged as the most consistent mechanistic correlate of CM-hAMSC activity.