INSTRUCTION: STEMIN-1

stemInduce™ MAG
Umbilical Cord Mesenchymal Stem Cell Assembly Medium

Description

The Intrinsic properties of many types of cells are critically altered when cell culture condition shifts from two-dimensional (2D) to three-dimensional (3D) environment. Currently, several lines of evidence have demonstrated the therapeutic potential of mesenchymal stem cells (MSCs) in regenerative medicine. The advantages of 3D grown MSC include the promotion of self-renewal
and differentiation activities as well as up-regulation of paracrine secretion. Following the development of 3D cell culture, the enhanced therapeutic efficacy of spheroid-forming MSCs have been identified in several animal disease models by promoting differentiation or trophic factor secretion, as compared to planar-cultured MSCs.

Spheroids are produced using different techniques such as “hanging drop” or “low attachment” surfaces. These techniques rely on the cells attaching to each other by the lack of another alternative. So, in this scaffold-free platform, cells are organized into the cell spheroids and floated in the culture medium due to the lack of tangible supporting materials. In contrast to that, the medium stemInduce ™ MAG from SMSbiotech induces the active reorganization of umbilical cord mesenchymal stem cells that grown on a regular, cell-adherent plate. The cells migrate towards a center and organize into a complex 3D cell assembly. The initiation of this complex organization follows immediately after 2D cell confluency and continues for many days.

*Shelf life may vary based on date of manufacturing

Not provided but required: Antibiotics (recommended: 10 µl Gentamicin & 0.25 µg/ml Amphotericin B)

Product use

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 Supplement MAG (50X) in a non-frost-free freezer. Avoid multiple freeze/thaw cycles.
• Do not freeze Basal medium MAG.

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.

Prepare Supplement MAG Supplemented Basal medium MAG
1. Thaw the SMAG (50X) solution in a 37°C water bath or overnight at 4°C. If thawed in a water bath, do not leave the vial at 37°C after the solution has thawed.
2. Aseptically transfer the contents of the thawed SMAG-1-1 solution to a bottle containing BMAG-1-100. Tightly cap the bottle and swirl to mix. Avoid causing medium to foam.
3. Store supplemented medium in the dark at 4-7°C. Do not freeze supplemented medium. When stored properly, stemInduce™ MAG medium is stable for up to 3 weeks.

UC-MSC assembly formation assay:

The following procedure is designed to induce spontaneous self-assembly of umbilical cord (Wharton jelly) derived mesenchymal stem cells:
1. Equilibrate a sufficient volume of supplemented stemInduce™ MAG at 37°C in humidified atmosphere containing 5% CO2.
2. Rapidly thaw cryopreserved UC-MSCs 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.
3. Add appropriate amount of UC-MSCs to plate or flask containing either UC-MSC stemInduce™ MAG medium.
4. Replace stemInduce™ MAG medium with similar amount of equilibrated supplemented stemInduce™ MAG medium the next day.
5. Replace stemInduce™ MAG medium with similar amount of equilibrated supplemented stemInduce™ MAG medium every 2-3 days until confluency.
6. To minimize disturbance to assemblies, we recommend replacing only half the amount of equilibrated supplemented stemInduce™ MAG medium every 2-3 days, after confluency.

Expected results
• Umbilical Cord MSC start assembling right after confluency and may form several assemblies, or even a singlelarge one.
• The process is relatively fast and requires few days.
• some assemblies may detach from the bottom.

Limited product warranty
SMSbiotech, inc. warrant 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.