StemXVivo GMP Mesenchymal Stem Cell Expansion Media
StemXVivo GMP Mesenchymal Stem Cell Expansion Media Summary
Complete, fetal bovine serum-containing media to support the expansion of mesenchymal stem cells (MSCs) under GMP-grade culture conditions.
- Produced according to GMP Guidelines
- All components, including fetal bovine serum, were selected and optimized for MSCs
- Formulated to reduce variation
- Ready to use
- Additional testing and documentation is available upon request
Why Culture MSCs GMP-grade Medium?
Mesenchymal stem cells (MSC) hold significant promise for tissue engineering and regenerative medicine. The use of MSCs for translational research requires their expansion in GMP-grade media and, depending on the application, GMP-grade cytokines and growth factors. GMP-grade media and proteins are manufactured so that all products, materials, equipment, and processes used in production are traceable and controlled. This extra level of traceability is important and necessary for transitioning MSC research into a translational, FDA-approved therapy.
Our StemXVivo® GMP Mesenchymal Stem Cell Expansion Media is the same formulation as our StemXVivo® Mesenchymal Expansion Media (Catalog # CCM004), but is produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.
StemXVivo® GMP Mesenchymal Stem Cell Expansion Media:
- Supports reproducible MSC growth in vitro.
- Can be supplemented with user-defined GMP-grade cytokines and growth factors, although these are not needed for culture.
StemXVivo® GMP Mesenchymal Stem Cell Expansion Media is supplied in a 250 mL volume and contains high quality factors to support MSC expansion in vitro. Contains fetal bovine serum.
Stability and Storage
Store unopened reagents in the dark at < -20 °C in a manual defrost freezer. Do not use past the expiration date.
When handling biohazardous materials such as human cells, safe laboratory procedures should be followed and protective clothing should be worn.
- 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.
- This reagent should not be used beyond the expiration date indicated on the label.
- Results may vary due to variations among cells derived from different donors.
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Figure 1: Phenotypic Analysis of Human MSCs Expanded in MSC Expansion Media. Human MSCs were expanded using StemXVivo® GMP MSC Expansion Media. Filled histograms indicate cells stained with markers of undifferentiated MSCs including anti-CD105 (R&D Systems, Catalog # FAB1097A), anti-CD73 (R&D Systems, Catalog # FAB5795P), anti-CD90 (R&D Systems, Catalog # FAB2067A), or anti-CD45 (R&D Systems, Catalog # FAB1430P). The open histograms show isotype-matched control staining. MSCs appropriately express high levels of CD105, CD73, and CD90, and lack expression of CD45.
R&D Systems' GMP media is produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.
R&D Systems' quality focus includes:
- Manufacturing and testing under an ISO 9001:2008 and ISO 13485:2003 certified quality system
- Documented processes and QA control of documentation and process changes
- Personnel training programs • Raw material testing and vendor qualification/monitoring
- Fully validated equipment, processes and test methods
- Equipment calibration schedules using a computerized calibration program
- Facility maintenance, safety programs and pest control
- Material review process for variances
- Monitoring of stability over product shelf life
Additional testing and documentation requested by the customer can be arranged at an additional cost.
Production records and facilities are available for examination by appropriate personnel on site at R&D Systems in Minneapolis, Minnesota USA. R&D Systems sells its GMP grade media products for research use or further manufacturing use in ex vivo cell therapy applications. They are not for in vivo use or for use as therapeutic or other drugs, biologic products or devices. Please read the following End User Terms prior to using this product.
The End User is aware that R&D Systems, Inc. sells its GMP products for research use only or further manufacturing and not for in vivo use, the production of therapeutics or other drugs or for biologic products or devices. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.
Preparation and Storage
Background: Mesenchymal Stem Cells
The term 'mesenchymal stem cells' (MSCs) is most commonly used to describe multipotent self-renewing cells that can be differentiated in vitro to generate adipocytes, chondrocytes, and osteoblasts. However, because these biological properties and hierarchical relationships remain to be clearly demonstrated in vivo, the term 'multipotent mesenchymal stromal cells' is often used to distinguish cultured cells from their in vivo precursors. Originally discovered in mouse bone marrow, multipotent mesenchymal stromal cells cultured from a variety of species and tissue types, have been shown to differentiate into progeny of additional lineages including, cardiomyocytes, endothelial cells, hepatocytes, and neural cells. Again, the physiological relevance of these findings remains to be determined.
Refer to the product datasheet for complete product details.
Briefly, MSCs are cultured in the GMP MSC Expansion Media using the following protocol:
- Plate MSCs in expansion media
- Culture MSCs to 80-90% confluency
- Passage MSCs using fresh expansion media
Reagents supplied in the StemXVivo® GMP Mesenchymal Stem Cell Expansion Media (Catalog # CCM026):
- 250 mL of GMP-grade MSC expansion media
- Bone marrow-derived MSCs
- Penicillin-Streptomycin (100X)
- Trypsin-EDTA (10X)
- Phosphate-Buffered Saline (PBS) (Tocris; Catalog # 3156)
- 75 cm2 tissue culture (T75) flasks
- 15 mL centrifuge tubes
- Serological pipette
- Pipettes and pipette tips
- 37 °C and 5% CO2 incubator
- Centrifuge (low speed clinical or equivalent)
- Inverted Microscope
- Water Bath
This protocol has been tested using bone marrow-derived MSCs. If using a different tissue source or cell line, the protocol below may need to be modified.
StemXVivo® GMP Mesenchymal Stem Cell Expansion Media - Thaw the MSC Expansion Media at 2-8 °C or room temperature.
Completed StemXVivo® GMP MSC Expansion Media - Add Penicillin-Streptomycin (100X) to the StemXVivo® GMP MSC Expansion Media at a 1:100 dilution. Note: If Penicillin-Streptomycin is not needed for the experiment, it can be omitted.
Trypsin-EDTA (1X) - Dilute Trypsin-EDTA (10X) to 1X using PBS.
Culturing Mesenchymal Stem Cells
Add 3.5 - 4.0 x 105 MSCs resuspended in 20 mL of the pre-warmed StemXVivo® GMP MSC Expansion Media to a T75 flask.
Replace the medium every 3 days with fresh Complete StemXVivo® GMP MSC Expansion Media.
Culture the MSCs to 80-90% confluency.
Subculturing Mesenchymal Stem Cells
Remove and discard the media from the T75 tissue culture flask(s) containing the MSCs.
Wash the cells with PBS.
Add 1-2 mL of pre-warmed Trypsin-EDTA to the MSCs and incubate the flask at 37 °C.
Add 5mL of Complete StemXVivo® GMP MSC Expansion Media to the flask to neutralize the Trypsin-EDTA.
Transfer the dissociated MSCs to a 15 mL conical tube.
Centrifuge at 400 x g for 5 minutes.
Resuspend the cell pellet in 5 mL of StemXVivo® GMP MSC Expansion Media.
Perform a cell count.
Add 3.5-4.0 x 105 MSCs resuspended in 20 mL of pre-warmed Complete StemXVivo® GMP MSC Expansion Media to each T75 flask.
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