STEMmatrix? BME
Basement membrane extract that supports feeder-free expansion and differentiation of hPSCs
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Overview
STEMmatrix? BME is a soluble, hPSC-qualified, basement membrane matrix extracted from mouse Engelbreth-Holm-Swarm (EHS) sarcoma. Rich in key extracellular matrix (ECM) proteins (e.g. collagen IV, entactin, heparan sulphate proteoglycans, laminin) and essential growth factors (e.g. EGF, bFGF, IGF-1, TGF-β, VEGF), this ECM-based hydrogel closely mimics the in vivo environment to support robust cell growth. When used with feeder-free hPSC maintenance media, such as mTeSR? Plus, mTeSR?1, TeSR?-E8?, or eTeSR?, STEMmatrix? BME can be used to successfully maintain hPSC lines in the undifferentiated state. These cells retain characteristic hPSC morphology, express undifferentiated cell markers such as OCT4 and TRA-1-60, and have the capacity to differentiate into all three germ layers.
Access our online protocol for step-by-step instructions on how to store, thaw, dilute, and coat plates with STEMmatrix? BME for hPSC culture.
Data Figures
Figure 1. Robust hPSC Expansion Is Observed in STEMmatrix? BME Cultures, with Comparable Performance to Corning? Matrigel?
Average fold expansion of clump-passaged hPSCs (± SEM) over 10 passages was similar between cultures grown on STEMmatrix? BME and Corning? Matrigel? hESC-Qualified Matrix. Cultures were maintained in mTeSR? Plus (Catalog #100-0276) or mTeSR?1 (Catalog #85850) media at 37°C and passaged every 7 days. Graph shows pooled data from H9 and SCTi003-A (Catalog #200-0511) hPSC cultures.
Figure 2. STEMmatrix? BME Supports Large hPSC Colony Sizes, with Comparable Performance to Corning? Matrigel?
Colony sizes of H9 and SCTi003-A (Catalog #200-0511) hPSCs grown on STEMmatrix? BME in mTeSR? Plus (Catalog #100-0276) or mTeSR?1 (Catalog #85850) media (± SD), averaged over 10 passages, were comparable to those grown on Corning? Matrigel? hESC-Qualified Matrix.
Figure 3. Normal hPSC Morphology Is Observed in Cells Cultured on STEMmatrix? BME
Microscope images are shown of (A) H9 and (B) SCTi003-A (Catalog #200-0511) PSCs grown on STEMmatrix? BME or Corning? Matrigel? hESC-Qualified Matrix in either mTeSR? Plus (Catalog #100-0276) or mTeSR?1 (Catalog #85850) media, all o f which exhibit comparable, normal morphology. Cells were imaged on the day of passage (Day 7) and display a highly multilayered and densely packed appearance.
Figure 4. hiPSC Morphology Is Consistent Across STEMmatrix? BME Lots and Comparable to Corning? Matrigel? Control
Representative images show WLS-1C hiPSCs cultured in mTeSR? Plus (Catalog #100-0276) on Corning? Matrigel? hESC-Qualified Matrix or one of three STEMmatrix? BME lots. Morphology was normal and consistent across all STEMmatrix? BME lots tested, and comparable to the Matrigel? control.
Figure 5. Undifferentiated Cell Markers Are Highly Expressed in hPSCs Grown on STEMmatrix? BME, with Comparable Expression Levels to Corning? Matrigel?
H9 and SCTi003-A (Catalog #200-0511) hPSCs were cultured in mTeSR? Plus (Catalog #100-0276) or mTeSR?1 media (Catalog #85850) with either STEMmatrix? BME or Corning? Matrigel? hESC-Qualified Matrix, then characterized for undifferentiated cell markers using flow cytometry. Within each cell and media pairing, STEMmatrix? BME and Corning? Matrigel? conditions exhibited similar expression levels of the undifferentiated cell markers OCT4 (A) and TRA-1-60 (B). Graphs show the average gene expression (± SD) from duplicate wells every 5 passages, up to a total of 10 passages.
Figure 6. hPSCs Grown on STEMmatrix? BME and Differentiated into Mesoderm, Endoderm, and Ectoderm Cells Show High Expression of Germ Line-Specific Markers, with Results Comparable to Corning? Matrigel?
Fold expression changes of mesoderm-, endoderm-, and ectoderm-specific genes are shown for cells derived from the SCTi003-A hiPSC line (Catalog #200-0511) that were grown on either STEMmatrix? BME or Corning? Matrigel? in mTeSR? Plus (Catalog #100-0276). Cells were grown and differentiated using the STEMdiff? Trilineage Differentiation Kit (Catalog #05230) and STEMdiff? SMADi Neural Induction Kit (Catalog #08581) on each respective matrix, and harvested once confluent. qPCR analysis was completed using the Human Pluripotent Stem Cell Trilineage Differentiation qPCR Array (Catalog #07515) and 海角破解版’s online qPCR analysis tool. For additional information about the trilineage gene expression markers included in this evaluation, or to explore the qPCR analysis tool, visit .
Figure 7. hiPSC-derived Mesoderm, Endoderm, and Ectoderm Cells Grown on STEMmatrix? BME Demonstrate High Expression of Germ Line-Specific Markers in Flow Cytometry Analysis
WLS-1C hiPSCs were grown on STEMmatrix? BME or Corning? Matrigel? hESC-Qualified Matrix in either (A) mTeSR? Plus (Catalog #100-0276) or (B) mTeSR? 1 (Catalog #85850), then dif ferentiated to mesoderm, endoderm, and ectoderm cells using the STEMdiff? Trilineage Differentiation Kit (Catalog #07515) on each respective matrix. Once confluent, cells were harvested, and expression of germ line-specific markers was evaluated using flow cytometry. The mesoderm, endoderm, and ectoderm-specific markers were highly expressed in each respective germ line, and expression levels (± SD) were comparable between STEMmatrix? BME and Corning? Matrigel? conditions.
Figure 8. hPSCs Cultured on STEMmatrix? BME Are Karyotypically Normal
A genetic analysis of H9 hPSCs grown on STEMmatrix? BME or Corning? Matrigel? hESC-Qualified Matrix in mTeSR?1 medium (Catalog #85850) was performed using the hPSC Genetic Analysis Kit (Catalog #07550). No karyotypic abnormalities were observed in either STEMmatrix? BME or Corning? Matrigel? conditions. Data are reported as fold expression change relative to a genomic DNA control (± SD).
Figure 9. hPSCs Grown on STEMmatrix? BME Exhibit Robust hPSC Expansion, High Expression of Undifferentiated Markers, and Normal Morphology, with Comparable Results to Corning? Matrigel?
H9 hPSCs were expanded in single-cell culture on either STEMmatrix? BME or Corning? Matrigel? hESC-Qualified Matrix in eTeSR? maintenance medium (Catalog #100-1215). Cultures were incubated at 37°C and passaged every 4 or 5 days. (A) Average fold expansion per day (±SD), calculated over 10 passages, was similar between the two matrices. (B) Accumulated cell numbers (±SD), shown over 10 passages, were also comparable between the two matrices. (C) Expression levels of OCT4 and TRA-1-60 (±SD), markers of the undifferentiated state, were similar between STEMmatrix? BME and Corning? Matrigel? conditions. Bars represent the average of duplicate wells every 5 passages, up to a total of 10 passages. (D) hPSCs grown on STEMmatrix? BME or Corning? Matrigel? demonstrated normal, comparable morphology in microscope images. Cells were imaged on the day of passage (Day 4 or 5) and appeared densely packed.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
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Resources and Publications
Educational Materials (7)
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STEMmatrix? BME
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.