Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract

Figure 1. Goblet Cells in Adult Stem Cell-derived Human Descending Colon Organoids Adult stem cells isolated from human descending colon were embedded in Cultrex UltiMatrix RGF BME and cultured in growth medium for 30 days. Organoids were fixed and stained with MUC2 (green; Catalog # NBP2-44431), to visualize intestinal goblet cells, and counterstained for E-Cadherin (red; Catalog # AF748) and DAPI (blue; Catalog # 5748).

Figure 2. Cultrex UltiMatrix shows improved lot-to-lot consistency with low endotoxin and high protein concentration across 20 recent lots of Cultrex UltiMatrix. The brown lines represents the endotoxin levels measured in 20 recent lots of Cultrex UltiMatrix. The graph shows consistently low levels of endotoxin across the lots. Cultrex UltiMatrix is specifically designed to maintain low endotoxin levels. The blue lines represents the protein concentration measured in the same 20 lots of Cultrex UltiMatrix. The graph demonstrates the consistency of protein concentration from lot to lot, highlighting the product's ability to maintain higher, constant protein levels. Cultrex UltiMatrix is designed to provide consistent protein concentration for reliable experimental outcomes.

Figure 3. Cultrex UltiMatrix RGF Basement Membrane Extract (BME) Displays Consistent Lot-to-Lot ECM Composition, High Tensile Strength, and Gel Rates A) Simple Western analysis for Entactin, Laminin, and Collagen IV across two lots of both Cultrex UltiMatrix RGF BME (R&D Systems, Catalog # BME001) and an alternate commercial BME matrix. Cultrex UltiMatrix RGF BME shows consistent expression of Entactin, compared to the alternate ECM matrix. B) The tensile strength (elastic modulus) dynamics across two lots of Cultrex UltiMatrix RGF BME and an alternate commercial BME matrix. The elastic modulus was measured at increasing temperatures to determine tensile strength and gelling rate across two independent lots of Cultrex UltiMatrix RGF BME (R&D Systems, Catalog # BME001) and the alternate BME matrix. Cultrex UltiMatrix RGF BME was found to display consistent gelling rates and high tensile strength (storage modulus) compared to the alternate commercial BME matrix.

Figure 4. Human Descending Colon Organoids Grown in Cultrex UltiMatrix RGF BME. Adult stem cells isolated from human descending colon were embedded in Cultrex UltiMatrix RGF BME and cultured in growth medium for 30 days prior to imaging. A) Brightfield image of descending colon organoid showing tissue invagination and epithelial layer formation. B, C) Descending colon organoids were stained with Chromogranin A (green; Catalog # MAB90981), to visualize intestinal enteroendocrine cells, and counterstained for E-Cadherin (red; Catalog # AF748) and DAPI (blue; Catalog # 5748).

Figure 5. Human Lung Organoids Grown in Cultrex UltiMatrix RGF BME. Adult stem cells isolated from human lung biopsy tissue were embedded in Cultrex UltiMatrix RGF BME and cultured in lung organoid expansion medium where they were able to differentiate and exhibit key markers for various lung cell types. A) Lung organoids stained with anti-Cytokeratin 5 (KRT5) (green; Novus, Catalog # NB110-56916) and with anti-p63/ TP73L (red; R&D Systems, Catalog # AF1916) to visualize basal cells. B) Lung organoids stained with anti-Podoplanin (PDPN) (green; Novus, Catalog # NB600-1015) to visualize alveolar type I cells. C) Quantification of lung organoid size across organoids grown in Cultrex UltiMatrix RGF BME from 4 different lots shows high performance consistency of the matrix in this application.

Figure 6. Robust Induced Pluripotent Stem Cell (iPSC) Expansion Compared to an Alternate BME Supplier. (A) Human iPSCs were passaged in 6 well plates (seeded at 7.5x10⁵ cells/well) using either Cultrex UltiMatrix RGF BME (purple) or an Alternate BME Supplier (grey) as a substrate. At each passage cells were harvested and counted by hemocytometer. Cultrex UltiMatrix BME showed robust iPSC expansion compared to the Alternate BME Supplier. B) iPSC cells grown on Cultrex UltiMatrix BME retained expression of the pluripotency marker SSEA-4, as labeled using a Fluorescein conjugated SSEA-4 antibody (Catalog # FAB1435).

Figure 7. Improved Spheroid (Acinar) Maturation and Consistency Compared to Other Commercial Matrices Single MCF10A cells were cultured for in Cultrex UltiMatrix BME, Cultrex BME, Type R1 (3433-005-R1), or an alternate BME supplier. Over 12 days in culture, cells differentiated into luminal structures called “acini” and were quantified for acinar area. Cultrex UltiMatrix BME showed increased acinar maturation (larger area) compared to Cultrex BME, Type R1 and more consistent acinar area compared to the alternate BME supplier.

Figure 8. Optimized Extracellular Matrix Composition and Soluble Protein. A) Multiple lots of Cultrex UltiMatrix RGF BME were run by Simple Western to determine extracellular matrix protein composition. The Western blots show the matrix to have high lot-to-lot consistency of protein composition. B) Cultrex UltiMatrix RGF BME showed a lower amount of aggregated protein in the matrix solution compared to Cultrex BME, Type R1. Lower protein aggregation indicates a higher amount of soluble protein in solution, which improves protein crosslinking and matrix integrity in culture.