MimEX Irradiated Fibroblast Kit
MimEX Irradiated Fibroblast Kit Summary
MimEX™ Irradiated Fibroblasts and Media provide a feeder layer for the culture of adult epithelial stem cells.
Key Benefits
- Irradiated Fibroblasts promote adult stem cell isolation and expansion
- Kit provides enough cells to culture two 6-well plates
- Features ready-to-use media for culturing irradiated fibroblasts
- Cells are tested for their ability to support expansion of adult intestinal stem cells in vitro
- Quality control tested for consistent media performance
The Benefits of MimEX™ GI Differentiation Media for Generating 3-D Gastrointestinal Tissue In Vitro
The MimEX™ Tissue Model System allows for the expansion and differentiation of ground-state, adult stem cell populations. These cells create sustainable and accessible 3-dimensional tissues (3-D) in vitro and provide a valuable tool for biomarker discovery, disease modeling, drug screening*, and developmental biology studies. The MimEX™ Irradiated Fibroblast Kit component of this system supports the expansion and differentiation of adult epithelial stem cell populations under clonal conditions while maintaining regional identity, genome stability, and disease state.
The MimEX™ Irradiated Fibroblast Kit contains 6 vials of cryopreserved irradiated fibroblasts along with culture media and basement membrane extract. The cells and media have been tested for their ability to support expansion of adult intestinal stem cells in vitro.
This kit contains the following reagents to support adult epithelial stem cell expansion and differentiation:
- 6 vials of MimEX™ Irradiated Fibroblasts
- 50 mL of MimEX™ Fibroblast Media
- 1 mL of Cultrex® Stem Cell Qualified RGF Basement Membrane Extract
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.
 View Larger Image |
Irradiated Fibroblast Plating Density and Distribution the Day After Plating for Stem Cell Expansion. MimEX™ Irradiated Fibroblasts were plated and cultured for one day in MimEX™ Fibroblast Media. Seeding density was evaluated with phase contrast imaging which shows a dense monolayer of cells that are evenly distributed on the culture surface. |
* 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.
All trademarks and registered trademarks are the property of their respective owners.
Specifications
Product Datasheets
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, MimEX™ Irradiated Fibroblast Kit can be used to culture adult epithelial stem cell using the following procedure:
- Thaw MimEX™ Irradiated Fibroblast Media
- Coat a 6-well plate with Cultrex® Stem Cell-Qualified RGF BME
- Thaw cryovial and plate MimEX™ Irradiated Fibroblasts into a 6-well plate
- After 24 hours, fibroblasts are ready to support isolation, expansion, and differentiation of adult epithelial stem cells.
Reagents provided in the MimEX™ Irradiated Fibroblast Kit (Catalog # MIM005):
- 6 vials of MimEX™ Irradiated Fibroblasts
- 50 mL of MimEX™ Fibroblast Media
- 1 mL of Cultrex® Stem Cell Qualified RGF Basement Membrane Extract
Reagents
- DMEM/F12
- Penicillin-Streptomycin (100X), optional
Materials
- 6-well tissue culture treated plates
- 15 and 50 mL centrifuge tubes
- Serological pipettes
- Pipette and pipette tips
Equipment
- 37 °C and 7.5% CO2 incubator
- Inverted microscope
- water bath
The protocol below describes the plating of MimEX™ Irradiated Fibroblasts for the expansion or differentiation of epithelial stem cells.
Note: The detailed protocol must be read in its entirety before using this product.
Protocol to Prepare 6-well Plates for GI Stem Cell Expansion
Protocol to Prepare Transwell Inserts for GI Stem Cell Differentiation
Plating MimEX™ Irradiated Fibroblasts for Stem Cell Expansion in a 6-well Plate
The following protocol is to plate one vial of MimEX™ Irradiated Fibroblasts into a 6-well plate. If an alternate plate size is desired, adjust volumes and cell numbers accordingly.
Place a 6-well plate at 2-8 °C for 10 minutes.
Dilute thawed Cultrex® Stem Cell Qualified RGF BME 1:5 in ice cold DMEM/F12. Keep on ice.
Coat desired number of wells with diluted Cultrex® BME mixture by pipetting 1 mL into the center of a well, swirl to coat the entire well, and remove by pipette while tilting the plate. Repeat for all wells need.
Remove excess Cultrex® BME in the wells.
Incubate the plate at 37 °C/7.5% CO2 for 10 minutes.
Thaw MimEX™ Irradiated Fibroblasts in a 37°C water bath.
Add 0.5 mL of MimEX™ Fibroblast Media to the cryovial of fibroblasts and gently pipette to re-suspend.
Add the resuspended cells dropwise to 2 mL of MimEX™ Fibroblast Media in a 50 mL centrifuge tube while swirling gently.
Add the fibroblast suspension to the coated wells, 2 mL/well. Ensure there are no air bubbles in the wells.
Distribute cells in the well using the MimEX™ Cell Plating Procedure. (Critical Step)
Incubate the plate in a 37 °C/7.5% CO2 incubator. (Critical Step) without disturbing the plate.
Replace MimEX™ Fibroblast Media daily. Plated fibroblasts should be used within 1-3 days of plating.
Plating MimEX™ Irradiated Fibroblasts for Stem Cell Differentiation in a 24-well Transwell Insert Plate.
The following protocol is for 12 transwell inserts of a 24-well plate. If an alternate plate size is desired, adjust volumes and cell numbers accordingly.
Place a 24-well plate with the desired number of transwell inserts at 2-8 °C for 10 minutes.
Dilute thawed Cultrex® Stem Cell Qualified RGF BME 1:5 in ice cold DMEM/F12. Keep on ice.
Coat desired number of wells with diluted Cultrex® BME mixture by pipetting 100 µL into the center of a transwell insert, swirl to coat the entire transwell insert, and remove by pipette while tilting the plate. Repeat for all wells need.
Remove excess Cultrex® BME in the transwell inserts.
Incubate the plate at 37 °C/7.5% CO2 for 10 minutes.
Thaw MimEX™ Irradiated Fibroblats in a 37°C water bath.
Add 0.5 mL of MimEX™ Fibroblast Media to the cryovial of fibroblasts and gently pipette to re-suspend.
Add the cells dropwise to a 50 mL centrifuge containing MimEX™ Fibroblast Media (250 µL for each transwell used) while swirling gently.
Add the fibroblast suspension to the coated wells, 250 µL/well. Ensure there are no air bubbles in the wells.
Distribute cells in the well using the MimEX™ Cell Plating Procedure. (Critical Step)
Incubate the plate in a 37 °C/7.5% CO2 incubator.(Critical Step) without disturbing the plate.
Replace MimEX™ Fibroblast Media daily. Plated fibroblasts should be used within 1-3 days of plating.
FAQs
-
Does MIM005 contain Rho kinase inhibitors?
No.
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