Figure 1. STEMmatrix™ Spike-In Enables Fast, Flexible Matrix Addition

To use STEMmatrix™ Spike-In, prepare the culture matrix according to the Product Information Sheet (PIS). No thawing of the matrix or pre-coating of culture plates is needed. The matrix can be added after the medium is added to the plate, followed by cell seeding (Stepwise Addition Protocol) or, alternatively, prepared as a mixture of medium, matrix, and cells, then aliquoted into the culture vessel (Batch Protocol). This streamlined workflow reduces total plating time to less than 2.5 minutes, representing a significant time saving compared to traditional matrix coating methods for human pluripotent stem cell (hPSC) culture.

Figure 2. STEMmatrix™ Spike-In Promotes High-Quality hPSC Morphology

Human induced pluripotent stem cells (hiPSCs; SCTi003-A) and human embryonic stem cells (hESCs; H9) cultured in mTeSR™ Plus, TeSR™-AOF, or eTeSR™ media with STEMmatrix™ Spike-In exhibit morphology consistent with high-quality, undifferentiated growth. In mTeSR™ Plus and TeSR™-AOF, cells form compact colonies with defined, smooth edges, while eTeSR™ cultures display a uniform, homogeneous monolayer morphology. Colony morphology appears well-defined and uniform relative to typical Corning® Matrigel®-based cultures.

Figure 3. STEMmatrix™ Spike-In Supports High Levels of Expansion Comparable to Corning® Matrigel®

Fold expansion of hPSCs (mean ± SEM, n = 5) cultured with STEMmatrix™ Spike-In or Corning® Matrigel® using mTeSR™ Plus or TeSR™-AOF media. Expansion values for mTeSR™ Plus and TeSR™-AOF were measured over 7-day aggregate passages. Within each media condition, STEMmatrix™ Spike-In consistently supported expansion comparable to Corning® Matrigel®, demonstrating reliable performance across multiple culture systems.

Figure 4. hPSCs Maintained on STEMmatrix™ Spike-In Retain Efficient Tri-Lineage Differentiation Capacity

Human iPSCs (WLS-1C) and hESCs (H1) were maintained using STEMmatrix™ Spike-In prior to and during directed differentiation. Cells efficiently differentiated into (A) ectoderm (PAX6⁺/Nestin⁺), (B) mesoderm (Brachyury⁺/NCAM⁺), and (C) endoderm (CXCR4⁺/SOX17⁺) lineages using the STEMdiff™ SMADi Neural Induction Kit, STEMdiff™ Mesoderm Induction Medium, and STEMdiff™ Definitive Endoderm Kit, respectively, confirming preservation of pluripotency and developmental potential following culture. Representative lineage-specific differentiation outcomes include (D) forebrain neurons generated using the STEMdiff™ Forebrain Neuron Differentiation Kit and (E) cardiomyocytes generated using the STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit following transfer to Corning® Matrigel®, as well as (F) hepatic progenitor-like cells generated using the STEMdiff™ Hepatocyte Kit with STEMmatrix™ Spike-In.