INSTRUCTION: SCEM-1

sceMatrix™ 24-well plate

Description
The coated 24 well plate has been optimized for the culturing of various human primary cells? or cell lines. The plate contains a thin layer of the small mobile stem cell-derived extracellular matrix (sceMatrix™). The following protocol provides instructions for assay

*Only 20 wells are coated **Shelf life may vary based on date of manufacturing

Product use

Protected under U.S. Pat. No. 10041037B2; all SMSbiotech Inc. products are to be used for research purposes only, and for no other purpose. The phrase research purposes only means scientific research programs directly under the user’s control, which are specifically directed to the purposes of internal research and not for any Commercial Purpose.

Important information

• Store plates coated with SMS-ECM-1 in the Freezer (-20°C) in intact, vacuum-sealed, plastic pouch until ready for use.

Safety information

Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. Caution: Human origin materials are non-reactive (donor level) for anti-HIV 1 & 2, anti-HCV, and HBsAg. Handle in accordance with established bio-safety practices.

Cell incubation

The following procedure was designed for the seeding of cells onto a 24-well plate. Please note that the 4 wells in column 1 (A1 to D1) are not coated with SMS-ECM-1 and can serve as control wells if needed.
1. Remove the plate from the freezer and allow the plate to come to room temperature for 30-40 min.
2. Carefully remove the plastic cover inside of a class 2 biosafety hood.
3. Equilibrate a sufficient volume of medium without supplement to wash the coated wells by incubating the un-supplemented medium at 37°C in humidified atmosphere containing 5% CO2. Use approximately 35 ml medium for each 24-well plate, or 1.5 ml for each of the 20 coated wells.
4. Equilibrate a sufficient volume of medium with the supplement (+ antibiotics if needed) by incubating the supplemented medium at 37 C° in the humidified atmosphere of 5% CO2. Use approximately 13 ml supplemented medium for each 24-well plate, or 0.5 ml for each of the 20 coated wells.
5. Wash the wells three times with 0.5 ml equilibrated un-supplemented medium. Do not touch the bottom of the plate with the vacuum tip. Instead, you may tilt the plate and remove excess fluid from the side.
6. Add 0.5 ml equilibrated supplemented medium (+ antibiotics) to each well.
7. Incubate the 24-well plate at 37°C in a humidified atmosphere containing 5% CO2. The plate is now ready for the plating of cells.

Plating cells

1. Rapidly thaw cryopreserved in a 37°C water bath. Please ensure that the tube is incubated for less than 90 seconds and that the cell suspension is only marginally thawed. After 90 seconds in the water bath, mix the cells and continue to thaw them gently at room temperature.
2. Use 20 μl of the suspension to determine the amount and viability of the cells.
3. Add desired amount of the cell suspension to each well.
4. Cover the plate, and gently shake horizontally to disperse the cells within the well.

Incubating the 24-well plate

1. Incubate the 24-well plate at 37 °C in a humidified atmosphere containing 5% CO2.
2. The following day, replace the medium in each well with 0.5 ml of fresh supplemented medium (+ antibiotics). Do not touch the bottom of the plate with the vacuum tip. Instead, you may tilt the plate and remove excess fluid from the side.
3. repeat step 2 as desired.

Visualization of cells

Cells and tissues can be visualized using fluorescent or non-fluorescent dyes.
Expected results

Limited Product Warranty
SMSbiotech inc. warrants their products as set forth in the SMSbiotechs` General Terms and conditions of Sale found on
the website. If you have any questions, please contact us.

DISCLAIMER: SMSbiotech disclaim all warranties with respect to this document, expressed or implied, including but not limited to those of
merchantability, fitness for a particular purpose, or non-infringement. To the extent allowed by law, in no event shall SMSbiotech be liable,
whether in contract, tort, warranty, or under any statute or on any other basis for special, incidental, indirect, punitive, multiple or consequential
damages in connection with or arising from this document, including but not limited to the use thereof.