Human Mesenchymal Stem Cells - Amnion
General information
Description | Amnion-derived Human Mesenchymal Stem Cells (hMSCs) possess several distinctive features that differentiate them from MSCs derived from other tissues, such as bone marrow, adipose tissue, and umbilical cord. One of the most significant distinctions is their origin from the amnion, a membrane of the placenta, which endows them with unique biological properties. Unlike MSCs from adult tissues, amnion hMSCs are more primitive and exhibit higher proliferative capacity, allowing for extended expansion in culture without significant loss of differentiation potential or stemness. This high proliferative capacity is particularly advantageous for applications requiring large cell quantities, such as tissue engineering and regenerative medicine. Another key difference lies in the immunomodulatory properties of amnion hMSCs. These cells demonstrate enhanced immunosuppressive abilities compared to MSCs from other sources, making them highly effective in modulating immune responses. This property is especially useful in research focused on inflammatory diseases, autoimmune conditions, and graft-versus-host disease (GVHD). Amnion hMSCs also secrete a distinct profile of bioactive molecules, including anti-inflammatory cytokines and growth factors, which contribute to their superior capacity for promoting tissue repair and reducing inflammation in various in vitro models. Additionally, amnion hMSCs are known for their lower immunogenicity compared to MSCs derived from other tissues. This reduced potential to elicit an immune response makes them particularly suitable for allogeneic applications and co-culture systems, where interactions between different cell types are studied without the complication of immune rejection. Furthermore, amnion hMSCs are ethically sourced from the placental tissue of healthy donors, eliminating ethical concerns associated with MSCs derived from more invasive procedures, such as bone marrow aspiration. Collectively, these attributes make amnion hMSCs a unique and versatile tool for a wide range of biomedical research applications. |
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Organism | Human |
Tissue | Amnion |
Applications | Drug testing, regenerative medicine, disease research |
Characteristics
Age | Please inquire |
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Gender | Please inquire |
Ethnicity | Caucasian |
Morphology | Well-spread spindle shaped, fibroblast-like morphology for at least within 5 passages. Fewer than 2% cells exhibit spontaneous myofibroblast-like morphology within each passage. |
Cell type | Stem cell |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | Human Mesenchymal Stem Cells, Amnion (Cytion catalog number 300644) |
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Biosafety level | 1 |
Expression / Mutation
Antigen expression | A comprehensive panel of markers, including CD73/CD90/CD105 (positive) and CD14/CD34/CD45/HLA-DR (negative), are used in flow cytometry analysis to identify cultivated MSCs (P2-P3) prior to cryopreservation. These markers are recommended by the ISCT MSC committee. |
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Viruses | Donor is negative for HBV (PCR), Treponema pallidum (PCR), and HIV-1/2 (IFA). Cells are negative for HBV, HCV, HSV1, HSV2, CMV, EBV, HHV6, Toxoplasma gondii, Treponema pallidum, Chlamydia trachomatis, Ureaplasma urealyticum, and Ureaplasma parvum. |
Handling
Culture Medium | Alpha MEM, w: 2.0 mM stable Glutamine, w/o: Ribonucleosides, w/o: Deoxyribonucleosides, w: 1.0 mM Sodium pyruvate, w: 2.2g/L NaHCO3 |
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Medium supplements | Supplement the medium with 10% FBS, 2 ng/mL bFGF |
Passaging solution | Trypsin-EDTA |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Seeding density | 1 to 3 x 10^4 cells/cm^2 |
Fluid renewal | First fluid renewal after 24 hours, then every 2 to 3 days. |
Freeze medium | CM-1 (Cytion catalog number 800100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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Required products
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.