Human placental decellularized extracellular matrix improves pancreatic progenitor differentiation from human embryonic stem cells compared with defined extracellular matrix protein substrates

Cell replacement therapy for diabetes requires a scalable source of functional β-cells. The derivation of insulin-producing cells (IPCs) from human embryonic stem cells (hESCs) is a strategy to generate β-cells for this purpose. Cell therapy for diabetes requires a scalable source of functional β-cells. The derivation of insulin-producing cells (IPCs) from human embryonic stem cells (hESCs) is a strategy to generate β-cells for this purpose. Despite advances in differentiation protocols, the contribution of extracellular matrix (ECM) substrate complexity to the specification of hESCs into pancreatic progenitors (PPCs) has not been systematically characterized. This study evaluated human placenta-derived decellularized ECM (PL-dECM) against defined ECM protein substrates and Matrigel in directing hESCs toward PPCs. We hypothesized that the multi-component composition of PL-dECM would offer more effective niche-mimicking signals for pancreatic differentiation than simplified substrates. hESCs were differentiated into PPCs using a previously established protocol. Cells were cultured on four substrates: (1) Matrigel (control), (2) collagen IV, (3) collagen IV combined with laminin-521, and (4) PL-dECM. Differentiation efficacy was evaluated using quantitative real-time PCR (qRT-PCR) for pancreatic progenitor markers (PDX1, NKX6.1) and confirmed by immunofluorescence. Cell survival was assessed by qRT-PCR quantification of apoptosis-related gene expression (SURVIVIN, BAX, BCL2). PL-dECM supported PPC differentiation at levels comparable to Matrigel, significantly outperforming both collagen IV alone and collagen IV supplemented with laminin-521. Cells cultured on PL-dECM showed significantly higher expression of PPC markers, particularly PDX1, alongside a more favorable cell survival profile. The incorporation of laminin-521 into collagen IV did not improve differentiation, and the collagen IV/laminin-521 substrate remained markedly less effective than both PL-dECM and Matrigel. The current results suggest that PL-dECM is an effective xeno-free substrate for directing hESCs toward PPCs, with differentiation efficiency comparable to Matrigel and superior to defined ECM protein substrates. These findings are limited to the progenitor stage. Future research should characterize the bioactive constituents of PL-dECM, elucidate the mechanisms that underlying its differentiation poromoting effects, and determine whether it can support β-cell maturation and clinical translation.