The adipose tissue stem cell line RESSTORE01 (Master Cell Bank/Stock n°1—Donor RESSSTORE01, Batch n°: 591133643763) was cultured in the growth medium Alpha MEM (Gibco, Life technologies) supplemented with 5% human platelet lysate (Stemulate, Cook Medical, USA) and 1% Penicillin-Streptomycin (Lonza, Belgium). The medium was changed twice each week and cells were passaged when they reached 100% confluence. The cells were detached with TrypLE Select (Life Technologies™, Thermo Fisher Scientific) for 10 min at 37°C and then centrifuged at 1,000 rpm for 5 min. The RESSTORE01 cell phenotype was analyzed with flow cytometry (FACSAria Fusion Cell Sorter, BD Biosciences) at passage PX +1. Monoclonal antibodies against CD19-phycoerythrincyanine (PE-Cy7), CD45RO-allophycocyanin (APC), CD73-PE, CD90-APC (BD Biosciences), CD11a-APC, CD105-PE (R&D Systems Inc., Minneapolis, MN, USA), CD34-APC and HLA-DR-PE (Immunotools GmbH, Friesoythe, Germany) were used. The cells expressed (>95%) surface markers CD73, CD90, and CD105 and lacked the expression (<2%) of CD11a, CD19, CD34, CD45, and HLA-DR. Cells at passages PX +2/3 were used for the study.
Stemulate
Manufactured by Cook Medical
Sourced in United States
Stemulate is a laboratory equipment product designed for cell culture applications. It functions as a cell stimulation and activation device, enabling the manipulation and study of cellular behavior and responses. The core function of Stemulate is to provide a controlled environment for the in vitro cultivation and analysis of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
α mem, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Cd73 pe, by BD (3 mentions)
Facsaria fusion cell sorter, by BD (2 mentions)
Cd34 pe, by BD (2 mentions)
Cellstack, by Corning (2 mentions)
Cd105 apc, by BD (2 mentions)
Cd45 fitc, by BD (2 mentions)
Cd90 fitc, by BD (2 mentions)
Cellstack culture chambers, by Corning (2 mentions)
Recombinant trypsin, by Roche (2 mentions)
Se full blood count analyzer, by Corning (2 mentions)
Cd90 apc, by BD (1 mentions)
Tryple select, by Thermo Fisher Scientific (1 mentions)
Cd11a apc, by R&D Systems (1 mentions)
Cd105 pe, by R&D Systems (1 mentions)
Penicillin streptomycin, by Lonza (1 mentions)
Cd34 apc, by Immunotools (1 mentions)
Hla dr pe, by Immunotools (1 mentions)
Facscalibur analyser, by BD (1 mentions)
6 protocols using stemulate
1
Characterization of RESSTORE01 Adipose Stem Cells
2
Isolation and Characterization of hBM-MSCs
3
Isolation and Expansion of hMSCs from Pediatric Bone Marrow
4
ASC Influence on Neuronal Cells
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To study the effect of ASCs on neuronal cells, human ASCs (Master Cell Bank/Stock no. 1—Donor RESSTORE01, Batch no. 591133643763) were cultured in Alpha MEM (Gibco) supplemented with 5% human platelet lysate (Stemulate, Cook Medical) and 1% P/S. ASCs were isolated and cultured as previously described [26 (link), 27 (link)]. The ASC phenotype was analyzed by flow cytometry (FACSAria Fusion Cell Sorter, BD Biosciences) and was found to reflect a typical MSC immunophenotype featuring expression (>95%) of the surface markers CD73, CD90, and CD105 and no expression (<2%) of CD11a, CD19, CD34, CD45, and HLA-DR [26 (link)].
5
Isolation and Expansion of hMSCs
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Human samples were provided by the Imperial College Healthcare Tissue Bank (ICHTB, HTA license 12275) supported by the National Institute for Health Research Biomedical Research Centre at Imperial College Healthcare NHS Trust and Imperial College London. ICHTB is approved by the UK National Research Ethics Service to release human material for research (12/WA/0196). hMSC were generated from bone marrow aspirates (issued from sub-collection R16052) collected from the iliac crest of healthy pediatric donors with informed consent. The total number of nucleated cells was established with a Sysmex SE full blood count analyzer and then 10-25 × 106 cells/636 cm2 were plated in CellSTACK® culture chambers (Corning). Cells were cultured in alpha modified Eagle's medium, no nucleosides (αMEM, Gibco) supplemented with 5% human platelet lysate (Stemulate, Cook Medical) under standard culture conditions (37 °C in a humidified atmosphere of 5% CO2/95% air). After reaching 90–100% confluency (10–14 days), cells were detached with recombinant trypsin (Roche, DE) and re-seeded at 5000 cells/cm2. hMSC were expanded in basal culture medium consisting of αMEM with 10% fetal bovine serum (FBS, Gibco) until passage 7 and regularly checked by flow cytometry to confirm that they expressed CD90, CD105, and CD73 and were negative for CD34 and CD45 [20 ].
6
Isolation and Expansion of hBM-MSCs
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hBM‐MSCs were isolated from bone marrow aspirates collected from the iliac crest of healthy pediatric donors, with informed consent from their parents or guardians. Cells were seeded in CellSTACK (Corning, Sigma Aldrich, UK) culture chambers at 10–25 × 106/636 cm2 and cultured in αMEM supplemented with human platelet lysate (Stemulate, Cook Medical, USA). At 90%–100% confluency, cells were passaged and seeded at 5,000 cells per cm2. For immunophenotyping of hBM‐MSCs, the following antibodies were used in conjunction with a FACSCalibur analyzer (BD Biosciences, UK): CD90‐FITC, CD105‐APC, CD73‐PE, CD34‐PE, and CD45‐FITC (all from BD Biosciences). All human tissue was approved for use by the UK National Research Ethics Service (12/WA/0196) and was collected by the National Institute for Health Research, which is supported by the Imperial College Healthcare Tissue Bank (HTA license 12275). Cultures were found to express CD90, CD105, CD73 and not express hematopoietic markers CD34 and CD45 26 (data not shown). hBM‐MSCs were expanded in growth media (GM; αMEM + 10% fetal bovine serum [FBS], Thermo Fisher Scientific, UK) under standard conditions (5% CO2).
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