Reagents and instruments for immunology, cell biology and molecular biology.

Human Mesenchymal Stem Cells, Adipose Tissue

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Cat# 300645

Size : 1millioncells

Brand : CLS Cell Lines Service

Human Mesenchymal Stem Cells - Adipose Tissue

Description	MSCs, or multipotent mesenchymal stromal cells, are self-renewing multipotent cells that can differentiate into a wide variety of cell types. The in vitro direct differentiation of MSCs into at least three orthodoxal lineages-adipocytes, osteoblasts, and chondrocytes has been demonstrated. Using differentiation media, it is possible to differentiate cultivated MSCs into adipocytes, osteoblasts, and chondrocytes in vitro. Early passage cultured MSCs are cryopreserved using a specific cryomedium.?After thawing, each cryovial contains 1 x 10^6 (minimum 92% to 95% viability level by Trypan Blue dye exclusion test). The MSCs were collected from healthy donors which have given their informed consent for the donation of the cell material. Each batch of MSCs is subjected to strict quality control testing (both cell donors and cell cultures). The identification, purity, potency, viability, and appropriateness of cultured MSCs for the intended usage are all evaluated.
Organism	Human
Tissue	Adipose Tissue
Applications	Drug testing, regenerative medicine, disease research

Age	Please inquire
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

Citation	Human Mesenchymal Stem Cells, Adipose Tissue (Cytion catalog number 300645)
Biosafety level	1

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.
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.

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
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) or CM-ACF (Cytion catalog number 806100)
Handling of cryopreserved cultures	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Optionally, skip centrifugation but remove any remaining freezing medium after 24 hours. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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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