hMSC expansion medium consisted of mesenchymal stem cell growth medium (MesenPRO; Gibco), 100 U penicillin, 1000 U streptomycin and 2 mM l -glutamine (Gibco). Murine MSCs were cultured in low-glucose Dulbecco’s Modified Eagle’s medium (LGDMEM; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin-streptomycin-glutamine (PSG; Thermo Fisher Scientific). We used endothelial cell medium (ECM; Sigma-Aldrich) as a HUVEC culture medium and these cells were expanded on fibronectin-coated culture vessels.
Mesenpro
Manufactured by Thermo Fisher Scientific
The MesenPRO is a cell culture media formulated for the expansion and maintenance of human mesenchymal stem cells (hMSCs) in vitro. It provides a serum-free, xeno-free, and chemically defined environment to support the growth and differentiation of hMSCs.
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Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (4 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
α mem, by Thermo Fisher Scientific (2 mentions)
Dmem lg, by Merck Group (1 mentions)
Endothelial cell medium (ecm), by Merck Group (1 mentions)
Iscove modified dulbecco medium, by Thermo Fisher Scientific (1 mentions)
Mtt assay, by American Type Culture Collection (1 mentions)
Bm msc, by Lonza (1 mentions)
Admscs, by Thermo Fisher Scientific (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Foetal calf serum, by Thermo Fisher Scientific (1 mentions)
Mesenpro rs growth supplement, by Thermo Fisher Scientific (1 mentions)
α minimal essential medium, by Thermo Fisher Scientific (1 mentions)
5 protocols using mesenpro
1
Mesenchymal and Endothelial Cell Culture
2
Expansion Media for MSCs and Osteosarcoma Cells
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Expansion medium for MSCs consists of MSC growth medium (MesenPRO; Gibco), 100 U penicillin, 1,000 U streptomycin, and 2 mM l -glutamine (Gibco). Expansion medium for MG63 (osteoblast-like osteosarcoma cell line), SaOS2 and 143B (high-grade osteosarcoma cell line), and hFOB cells consists of Iscove modified Dulbecco medium (Gibco) and 10% fetal bovine serum (Hyclone, Logan, UT, USA) supplemented with 100 U penicillin, 1,000 U streptomycin, and 2 mM l -glutamine (Gibco).
3
Vitamin C Enhances hASC Proliferation
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Example 18
To evaluate the mitogenic effects of vitamin C and its derivatives on human adipose tissue derived stem cells (hASCs), hASCs were cultured on tissue culture plastic for 4 days in complete MesenPro medium (Invitrogen, Carlsbad, Calif.) supplemented with or without vitamin C (ascorbic acid) or its derivatives (Vitagen or AA2G) in free form. Proliferation was assessed by MTT assay as described by the manufacturer (ATCC, Manassas, Va.). After 4 days, concentrations of ascorbic acid, 0.25, 0.5, and 1 mM, were found to enhance proliferation (measured by amount of conversion of yellow tetrazolium MTT into purple formazan by dehydrogenase enzymes (the purple formazon is solubilized by detergent) by 60%, 80%, and 96% above controls lacking ascorbic acid, respectively. Using the same concentrations of AA2G yielded proliferation enhancements of 70%, 60%, and 50% above controls, respectively. Similar results were obtained with Vitagen, showing 70%, 60%, and 30% increases over controls, respectively. In summary, vitamin C and its derivatives, AA2G and vitagen, in the presence of growth factor containing media, enhance hASC proliferation in cell culture.
4
Expansion of Three MSC Types
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Three types of MSCs were expanded as previously reported [16 (link), 17 (link)]. BMMSCs (Cat no. PT-2501, Lonza, USA) were maintained in DMEM containing 10% FBS and 1% antibiotic-penicillin/streptomycin solution. ADMSCs (Cat no. R7788-115, Invitrogen) were expanded using MesenPRO RS™ basal medium, growth supplements and 2% FBS. SFMSCs were plated in complete culture medium consisting of α -MEM (Invitrogen) with 10% FBS, 1% antibiotic-penicillin/streptomycin solution and 250 ng/ml amphotericin B (Invitrogen). All types of cells were incubated at 37 °C under a humidified atmosphere containing 5% CO2.
5
Attachment of ADSCs on Ceramic Disks
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Human adipose-derived stem cells (ADSCs) were purchased from the Invitrogen Technologies and the cells from three donors were mixed and expanded to reach passage four by culturing in MesenPRO basal medium with 2 mM l -glutamine and MesenPRO RS Growth Supplement (Invitrogen). Passage 4 ADSCs were used for all the experiments. The attachment of ADSCs cultured for 24 h on the ceramic disks was analyzed using scanning electron microscopy. Briefly, the ADSCs were seeded on the disks at the density of 10,000 cells per mL and cultured for 24 h in growth medium consisting of α-minimal essential medium (α-MEM, Gibco Laboratories), supplemented with 10% (v/v) foetal calf serum (Gibco Laboratories), 100 U/mL penicillin, and 100 mg/mL streptomycin (Gibco Laboratories). After 24 h of culture, cells were rinsed in phosphate-buffered solution (PBS), fixed in 1.25% glutaraldehyde and sequentially dehydrated in graded ethanol. Samples were dried in hexamethyldisilazane and coated with gold for scanning electron microscopy (SEM) analysis using FE-scanning electron microscopy (Zeiss Ultra).
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