extracellular matrix
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Extragel Matrix Manual

Product Characteristics Operation Method Practical Application Cases Product Introduction

Product Introduction The basement membrane is a matrix under the basal surface of epithelial cells of animals. Scilia Extragel Matrix is a reconstituted matrix hydrogel formed by basement membrane components extracted from mouse tumor tissues. This matrix hydrogel is mainly composed of laminin, collagen IV, and heparan sulfate proteoglycans (Kleinman et al. 1986). Besides, it contains various growth factors, such as epidermal growth factor (EGF), platelet-derived growth factor (PDGF), nerve growth factor (NGF), basic fibroblast growth factor (FGF-2), transforming growth factor-β (TGF-β), and...

Operation Method ORGANOID CULTURE (1 HOUR) 1. Thaw the Extragel Matrix (Scilia-OM-1 or Scilia-OM-1-ph or Scilia-OM-3 or Scilia-OM-3-ph) in refrigerator at 4℃ overnight. 2. Preheat the 24-well plate in cell culture incubator. 3. Prepare aliquots of Extragel Matrix using pre-chilled tips. 4. Prepare the single cell pellet with 1×106 cells derived from patients or animal tissue, centrifuged at 300 g for 5 minutes. 5. Mix the cell pellet with 50 μL Extragel Matrix thoroughly. 6. Add the mixture into the well of plate (50 μL per well). 7. Keep the plate in incubator for 10 minutes, flip and keep...

Operation Method ANGIOGENESIS EXPERIMENT (1 HOUR) 1. Starving cells: replace the complement medium with starving medium: DMEM medium containing 0.2% FBS, 2 mM L-glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin, and 100 μg/mL streptomycin for the incubation for 24h. 2. Uniformly spread Extragel Matrix (Scilia-OM-1 or Scilia-OM-1-ph) on the 96-well plate. (Note: The pipette should be pre-cooled for 30 minutes. The operation shall be performed on ice to avoid the gelation of Extragel Matrix and the generation of bubbles.) 3. Incubate the 96-well plate in a cell incubator for 30 minutes to...

NEURITE OUTGROWTH ASSAY 1. Prepare the rat neural stem cell (NSC) pellet with 1×105 cells, centrifuged at 300 g for 5 minutes. 2. Mix the cell pellet with 50 μL Extragel Matrix (Scilia-OM-1 or Scilia-OM-1-ph), thoroughly. 3. Keep the plate in incubator for 10 minutes, flip and keep after another 5 minutes. 4. Add 500 μL culture medium to the well with matrix and cells. 5. Change the medium every 3 days. Notes: NSC culture medium contains Neurobasal medium, 2% B27, 2mM L-glutamine, 5% FBS, 20 ng/mL EGF, 20 ng/mL bFGF, 100 U/mL penicillin and 100 μ g/mL streptomycin. Neurites will be observed...

Practical Application Cases Figure 1. Establishment of human bile duct organoids in the matrix hydrogel of Company A and Extragel Matrix, respectively. Human bile duct organoids are imaged after being stained with anti-ZO-1 antibody (tight-junction protein), and Alexa Fluor 647 Phalloidin (cytoskeleton protein F-actin). Figure 2. Growth duct from human embryonic stem cells in the matrix hydrogel of Company A and

Practical Application Cases Figure 3. Growth of human colorectal cancer organoids derived from patients in the matrix hydrogel of Company A and Extragel Matrix for 13 days, respectively. Figure 4. Characterization of hepatocellular carcinoma organoids derived from patients in the matrix hydrogel of Company A and Extragel Matrix, with immunostaining by DAPI (nuclus), HNF4a (a hepatic biomarker) and GPC3 (hepatocellular carcinoma biomarker).

Practical Application Cases Company A Extragel Matrix Figure 5. Human umbilical vein endothelial cells (HUVECs) can form vascular networks in the matrix hydrogel of Company A and Extragel Matrix. HUVEC cells are imaged after being stained with living cell dye Calcein AM (green). Figure 6. Rat neural stem cells can grow, differentiate and form neurite networks in the Extragel Matrix. Rat neural stem cells (NSCs) were cultured in Extragel Matrix for 7 days and imaged after being stained with neuron biomarker Tuj1 (red) and nuclear dye DAPI (blue) (A). The right figure presents the neuron with...

Practical Application Cases Extragel Matrix Figure 7. Growth of hESCs on the matrix hydrogel of company A and Extragel Matrix for 2 days. Reference 1. Kleinman HK, et al, Basement membrane complexes with biological activity. Biochemistry 25: 312 (1986). 2. Vukicevic, Slobodan, et al. Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components. Experimental cell research 202: 1 (1992). 3. Guillen, K P, et al. A human breast cancer-derived xenograft and organoid platform for...

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