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- Other Names
- Mesenchymal Stromal Cells, Medicinal Signaling Cells, Multipotent Stromal Cells
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- Product Citations
- publications
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Figures 1. Human Bone Marrow-derived Mesenchymal Stem Cells (BM-MSCs) were seeded at a density of 6,000 cells/cm2 at 37°C and 5% CO2 in either αMEM containing 10% FBS or Cell-Vive™ MSC Xeno-Free Growth Media (cat#420519). Morphology of adherent cells grown in either (A) αMEM containing 10% FBS (B) or Cell-Vive™ MSC Xeno-Free Growth Media were observed 48-and 72-hours post-seeding. Cell expansion(C) and Cell viability (D) in αMEM containing10% FBS (white bar) or Cell-Vive™ MSC Xeno-Free Growth Media (cat#420519, black bar) were determined on Day 4. Representative BM-MSCs cultured in Cell-Vive™ MSC Xeno-Free Growth Media, GMP, on Day 5 were harvested & analyzed by Flow Cytometry (E) using PE anti-human CD105 (Clone 43A3, Cat#323206), PE/Cyanine7 anti-human CD90 (Clone 5E10, Cat# 328124), PE/Cyanine7 anti-human CD73 (clone AD2, Cat# 344044), FITC anti-human CD45 (Clone HI30, Cat# 304054), and APC anti-human CD34 (Clone 581, Cat#343510). Cells were positive for CD105, CD90 and CD73 but lacked CD45 and CD34 expression (Blue lined histogram). Isotype matched controls were included (Black lined histogram). -
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Figures 2. Human Adipocyte-derived Mesenchymal Stem Cells (AD-MSCs) were seeded at a density of 6,000 cells/cm at 37°Cand 5% CO2 in either αMEM containing 10% FBS or Cell-Vive™ MSC Xeno-Free Growth Media (cat#420519) for 5 days. Cell expansion (A) and cell viability (B) in αMEM containing 10% FBS (white bar) or Cell-Vive™ MSC Xeno-Free Growth Media (cat#420519, black bar) were determined on Day 5. Representative AD-MSCs cultured in Cell-Vive™ MSC Xeno-Free Growth Media on Day 5 were harvested and analyzed by Flow Cytometry (C) using PE anti-humanCD105 (Clone 43A3, Cat# 323206), PE/Cyanine7 anti-human CD90 (Clone 5E10,Cat# 328124), PE/Cyanine7 anti-humanCD73 (clone AD2, Cat# 344044), FITC anti-human CD45 (Clone HI30, Cat# 304054), and APC anti-human CD34 (Clone 581, Cat#343510). Cells were positive for CD105, CD90 and CD73 but lacked CD45 and CD34 expression (Green lined histogram). Isotype matched controls were included (Red lined histogram). -
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Figures 3. Human Bone Marrow-derived Mesenchymal Stem Cells (BM-MSCs) were cultured in Cell-Vive™ MSC Xeno-Free Growth Media (cat#420519). A cell proliferation assay was performed using CFSE-labeled human PBMC-derived CD3+ T cells stimulated with CD3/CD28 activation beads. Non-activated CD3+ T cells without MSC co-culture were arrested at the parent generation (Blue histogram). Activated CD3+ T cells proliferated for 4 days without MSC co-culture show low fluorescent intensity indicating cell division (Black histogram). Proliferation of CD3+ T cells stimulated with CD3/CD28 activation beads show higher fluorescent intensity when co-cultured with MSCs (Purple histogram), indicating less active CD3+ T cell proliferations from BM-MSCs-mediated immune modulations.
| Cat # | Size | Price | Quantity Check Availability | Save | ||
|---|---|---|---|---|---|---|
| 420519 | 250 mL | 220€ | ||||
| 420520 | 500 mL | 340€ | ||||
The Cell Vive™ MSC Xeno-Free Growth Media, GMP, is designed to support the expansion of human bone marrow-, adipose tissue- or umbilical cord tissue-derived mesenchymal stem cells (MSCs). Cell Vive™ MSC Xeno-Free Growth Media, GMP, is prepared without nonhuman components or phenol red. It is suitable for culturing and expansion of MSCs without the need to supplement with FBS. When used under appropriate conditions, this media demonstrates higher performance than FBS containing media without affecting the phenotype of the cultured cells. Performance is based on cell counts and viability. This GMP product is suggested for use in research and ex vivo cell processing. Benefits of this growth media:
- Xeno-Free, Serum-Free & without Phenol Red
- Additional supplementation with serums not required
- Able to support and maintain MSC expansion
- Manufactured in a GMP facility according to USP <1043>
BioLegend Cell-Vive™ GMP cell culture products are manufactured and tested in accordance with USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue- Engineered Products and Ph. Eur. Chapter 5.2.12 in a dedicated GMP facility compliant with ISO 13485:2016. Specifications and processes include
- Low endotoxin level (< 1 EU/mL)
- Mycoplasma and bacterial/fungal growth testing
- Batch-to-batch consistency
- Vendor qualification
- Raw material traceability and documentation
- Documented procedures and employee training
- Equipment maintenance and monitoring records
- Lot-specific certificates of analysis
- QA review of released products
- Quality audits per ISO 13485:2016
Product Details
- Formulation
- Xeno- and Serum-Free, GMP formulated MSC Xeno- Free Growth Media without Phenol Red.
- Endotoxin Level
- < 1 EU/mL
- Preparation
- Thaw media overnight or longer until fully thawed at 2-8°C. The media is ready to use after thaw.
- Storage & Handling
- Store at frozen between -20°C to -5°C. Thaw overnight at 2-8°C. If necessary, product can be kept at 2-8°C for up to two weeks.
- Application
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Cell Culture - Quality tested
Mesenchymal Stem Cell Culture - Verified in house
- Recommended Usage
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Cell-Vive™ MSC Xeno-Free Growth Media, GMP is a completed medium that is ready to use. Suggested cell seeding density of 6,000 cells/cm2 when used to support MSC expansion.
- Application Notes
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This product is a complete media formulation that is ready to use for culturing bone marrow-, adipose tissue- or umbilical cord tissue-derived mesenchymal stem cells. It can be used without any cell attachment. However, depending on the tissue sourced MSCs, recombinant human fibronectin (Cat#775314) can be used to further optimize the expansion.
The presence of trace precipitation in the product after thaw is normal and relates to the high enrichment of the product and should not impact its performance. The precipitates should go into solution after warming up the media in 37°C water bath or equivalent, or if desired, precipitates can be removed through 0.22 µm filtration of the media without impacting its performance.
Cell Culture Platform:
Cell-Vive™ MSC Xeno-Free Growth Media, GMP, can be used with tissue culture plates, and flasks. Further protocol optimization is recommended with the specific bioreactor being used.
MSC culture from cryopreserved human MSCs:
- When using tissue culture-treated culture ware, culture vessel may not need coating attachment substrate or use 5 µg/cm2 fibronectin-coated culture vessel.
- Pre-warm media to 37°C and avoid overheating.
- Rapidly thaw cryopreserved cells in the 37°C water bath.
- Remove the vial from the water bath once most of the solutions is thawed, the vial should remain cool to touch and a small ice fragment present.
- Wipe the external surface of medium bottle and vials with 70% ethanol and transfer to a biosafety cabinet.
- Resuspend cells using pre-warm media and seed MSCs at 6,000 cells/cm2 density.
- If cryopreservation solution contains DMSO. Extra steps needed.
- Carefully add 1 mL warm media dropwise to the cryovial while slowly stirring.
- Transfer all contents in cryovial to the conical tube and add 5-10 ml pre-warmed media dropwise into conical tube and gently stirring.
- Resuspend cells and seed MSCs at 6,000 cells/cm2 density.
- Incubate the cells at 37°C, 5% CO2. Aspirate off media and feed the cells with pre-warmed Cell-Vive™ MSC Xeno-Free Growth Media 24 hours after seeding.
- Every two days, aspirate media and feed the cells with pre-warmed Cell-Vive™ MSC Xeno-Free Growth Media.
- Subculture cells once they reach 80% confluence. Do not allow the culture to become overconfluent. Subculturing under suboptimal conditions may affect product performance.
Subculture MSCs
- Once cells reach 80 % confluency, passage cells and avoid overgrowth.
- Carefully aspirate the media from the culture vessel.
- Wash wells/flask with pre-warmed 1X PBS containing no calcium, no magnesium, no phenol red.
- Add TrypLE Express detachment solution or equivalent (Table 1) to cover the surface area of the well/flask.
- Incubate the cells in the 37°C and observe detachment in increments of 3-5 minutes.
- Once cells begin to detach, gently tap the culture vessel to dislodge cells. Repeat the process until at least 90% of cells are all detached.
- Stop cell dissociation by adding MSC Xeno-Free Growth Media at a 1:1 volume ratio.
- Transfer the cell suspension to a sterile conical tube.
- Centrifuge the cells at 300 x g for 5 minutes.
- Aspirate the media gently without disturbing the cell pellet.
- Resuspend cells using pre-warm media and seed MSCs at 6,000 cells/cm2 density.
Table 1. Suggested user guidelines for passaging
Vessel
Volume of PBS for wash (mL)
Volume of TrypLE Express (mL)
Incubation Time for Detachment (minutes)*
24 well
0.5
0.5
3 - 5
6 well
1
1
5 - 10
T-75
5
5
10 – 15
T-175
15
15
10 – 15
*incubation of detachment solution for cells may vary depending on the cell density, cell type, and level of adhesion to the plate. It is highly recommended to observe detachment in increments of 3-5 minutes and allow additional incubation if necessary.
- Disclaimer
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BioLegend Cell-Vive™ GMP Cell Culture products are for research use only. Suitable for ex vivo cell processing use. Not for injection or diagnostic or therapeutic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.
Antigen Details
- Gene ID
- NA
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