Liver Biomatrix Suspension is a novel new matrix isolated from decellularized liver tissue. The proprietary patent-pending process retains the liver matrix biochemistry leaving the matrix scaffold intact. Following the decellulariziation process, the matrix scaffold is reduced to um sized particles in suspension which is used for coating multi-well plates. Biomatrix coated plates provide the ultimate microenvironment for maintaining functional hepatocytes longer than when plated on collagen I. In addition, stem cells committed to the endodermal lineage differentiate into functional hepatocytes within a few days maintaining function for greater than 4 weeks.
Preparing Liver Biomatrix for Coating Plates:Liver Biomatrix suspension is easy to work with but requires adherence to coating instructions for successful coating of plates. Improper handling of the Biomatrix suspension can result in clumping of the biomatrix material and uneven coating of the plates. After coating plates with Biomatrix, it is essential for the plates to remain undisturbed for a minimum of 2 hours preferable overnight
1. Determine the number of plates you want to coat (see Table 1),2. Aseptically remove the plates from the packaging and place on the flat level surface in a biological safety cabinet or laminar flow hood. Organize the plates so that you can add Biomatrix suspension without moving the plates
3. Have pipettor, pipettes and reagent reservoir ready for use4. Thaw the vial of Liver Biomatrix Suspension by rapid thaw method in 37°C water-bath. Submerse the vial up to the cap in the water-bath and continually agitate in water until completely thawed –should take a couple minutes
a. Slow thawing can result in clumping of biomatrix material5. Coat plates immediately because allowing suspension to set for any length of time will result in clumping of the biomatrix material and uneven coating.
Coating Plates with Liver Biomatrix Suspension:1. Transfer appropriate volume of biomatrix suspension into each well using a multi-channel pipettor where appropriate. Refer to Table 1 for plating volume
a. Ensure that the suspension is evenly distributed across well – tap plate if necessary but don’t remove from flat surface of hood
2. Let plates remain undisturbed for a minimum of 4 hours allowing biomatrix material to settle to the bottom of the well. Best to coat plates first thing in the morning
a. This step is critical for even coating of the well with biomatrix particles
3. At the end of the day or after 4 hours remove solution from each wella. Handle plates carefully so matrix coating isn’t disturbedb. Tilt plate toward you without removing plate edge from hood surface. This stabilizes plate to prevent jerky movements that may make biomatrix come off the plate4. Using suction apparatus with fine tip pipettes aspirate solution from each well. DO NOT TOUCH MATRIX SURFACE WITH PIPETTE!!!a. Gently lay plate back down, leave lid off and allow to dry completely over night
i. Moving plate while wet disturbs the biomatrix coating which will result in an uneven coating of the well
5. When biomatrix coating is completely dry replace lid and examine for quality6. Coated plates may be stored in 4°C refrigerator for up to 1 year.
Storage and Stability:Store Liver Biomatrix Suspension Pack in -80°C freezer until ready to use
