MimEX GI Expansion Media

Kit Summary

Media for the expansion of adult human gastrointestinal stem cells.

The Benefits of MimEX™ GI Expansion Media for Generating 3-D Gastrointestinal In Vitro

The MimEX™ GI Tissue Model System allows for the expansion and differentiation of ground-state, adult stem cell populations from the gastrointestinal tract. These cells create sustainable and accessible 3-dimensional gastrointestinal tissues in vitro and provide a valuable tool for biomarker discovery, disease modeling, drug screening*, and developmental biology studies.

The MimEX™ GI Expansion Media component of this system will expand adult stem cell populations of the gastrointestinal tract under clonal conditions while maintaining regional identity, genome stability, and disease state. MimEX™ GI Expansion Media has been tested for its ability to support expansion of adult human intestinal stem cells in vitro when grown on plates coated with MimEX™ Irradiated Fibroblasts. When used in conjunction with MimEX™ Irradiated Fibroblasts, the expansion media will specifically select for and expand adult stem cells from a dissociated human tissue biopsy. These adult stem cells will retain local tissue memory, enabling the expansion and generation of in vitro organ tissue from specific regions of the gastrointestinal tract, including the ileum, jejunum, duodenum, and colon.

This kit contains the following reagents to isolate and expand adult gastrointestinal stem cells:

• 250 mL of MimEX™ GI Expansion Media

Precaution

When handling biohazardous materials such as human cells, safe laboratory procedures should be followed and protective clothing should be worn.

Limitations

• FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
• The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
• These reagents should not be used beyond the expiration date indicated on the label.
• Results may vary due to variations among adult stem cell lines originating from different donors.

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Intestinal Stem Cell Colony Morphology During Expansion. MimEX™ Descending Colon Stem Cells were cultured on fibroblast-coated plates in MimEX™ GI Expansion Media MimEX™ Descending Colon Stem Cells were cultured on fibroblast-coated plates in MimEX™ GI Expansion Media. Colony morphology was assessed using brightfield microscopy at 7 days of culture. Based on the size and distribution of the colonies, this culture is ready for passaging.

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Human Ascending Colon Adult Stem Cells Differentiate into Ascending Colon Tissue. Adult Human GI Stem Cells from the ascending colon were expanded on MimEX™ Irradiated Fibroblasts (R&D Systems®; Catalog # MIM005) in MimEX™ GI Expansion Media. They were then differentiated in MimEX™ GI Differentiation Media (R&D Systems®; Catalog # MIM004) for 9 days under air-liquid interface conditions. To evaluate differentiation, adult stem cell-derived ascending colon tissue was immersion-fixed, paraffin-embedded, and sections stained with Mouse Anti-Human Cadherin-17 Monoclonal Antibody (green; R&D Systems®; Catalog # MAB1032) and Rabbit Anti-Human/Mouse MUC2 Antibody (red; Novus Biologicals®; Catalog # NBP1-31231). Tissue was then stained using the NorthernLights™ (NL)493-conjugated Anti-Mouse IgG Secondary Antibody (R&D Systems®; Catalog # NL009) and NL557-conjugated Anti-Rabbit IgG Secondary Antibody (R&D Systems®; Catalog # NL004) and counterstained with DAPI (blue; Tocris®; 5748). Cadherin-17 staining was detected in cellular membranes throughout the epithelial layer while MUC2 staining was localized specifically to goblet cells.

* Portions of MimEX Tissue Model Systems are covered by a license to intellectual property owned by, or licensed to, Multiclonal Therapeutics, Inc. The purchase of this product conveys to the buyer the limited, non-exclusive, non-transferable right under these intellectual property rights to use this product for research purposes only. The purchase of this product does not include any right to use this product or components of this product for resale, for drug discovery intended for commercial gain, for the generation of cells or tissues intended for transplantation in human patients, or for any other commercial purposes, including without limitation, provision of services to a third party, generation of commercial databases, or as a diagnostic or therapeutic. Multiclonal Therapeutics, Inc., reserves all other rights under these rights. For information on purchasing a license to the intellectual property for uses other than in conjunction with this product or to use this product for purposes other than research, please contact Multiclonal Therapeutics, Inc. at licensing@multiclonaltherpaeutics.com.

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