T1-iMSCs and XSF-iMSCs (PN3–PN4) were stained on ice for 30 min with the following antibodies (1:50 dilution in FACS buffer): CD105-APC (eBioscience, 17-1057-42), CD90-PE (eBioscience, 555596), CD73-PE (eBioscience, 550257), CD44-PE (eBioscience, 550989), CD105APC (eBioscience, 17-1057-42), PE-isotype (BD Bioscience, 551438), and APC isotype (BD Bioscience, 565381). After washing, BD FACSAria™ III Cell Sorter was used to detect the fluorescence. The results were plotted using FlowJo_v10.8.1.
Cd105 apc
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Canada
CD105-APC is a flow cytometry reagent that specifically binds to the CD105 (Endoglin) antigen expressed on human cells. It is used for the identification and enumeration of CD105-positive cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Cd90 fitc, by Thermo Fisher Scientific (8 mentions)
Cd73 pe, by BD (8 mentions)
Cd73 pe, by Thermo Fisher Scientific (8 mentions)
Cd73 fitc, by Thermo Fisher Scientific (5 mentions)
Cd45 fitc, by Thermo Fisher Scientific (5 mentions)
Facsaria 3, by BD (4 mentions)
Facscalibur, by BD (4 mentions)
Cd29 fitc, by Thermo Fisher Scientific (4 mentions)
Accuri c6 flow cytometer, by BD (4 mentions)
Cd34 pe, by BD (4 mentions)
Cd45 pe, by Thermo Fisher Scientific (3 mentions)
Cd44 pe, by BD (3 mentions)
Cd45 fitc, by BD (3 mentions)
Cd31 fitc, by Thermo Fisher Scientific (3 mentions)
Hla dr fitc, by Thermo Fisher Scientific (3 mentions)
Cd34 percp, by Thermo Fisher Scientific (3 mentions)
Cd90 pe, by Thermo Fisher Scientific (3 mentions)
Cd44 pe, by Thermo Fisher Scientific (3 mentions)
Cd146 pe, by BD (3 mentions)
Cd34 fitc, by Thermo Fisher Scientific (3 mentions)
28 protocols using cd105 apc
1
Mesenchymal Stem Cell Characterization
2
Characterization of Mesenchymal Stem Cell Markers
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Cells were fixed with Cytofix/CytoPerm kit (BD Biosciences, USA) as per manufacturer’s instructions. Surface markers staining was performed using CD106-PE, CD271-PE, CD34-APC, CD105-APC, CD90-FITC, CD73-FITC and CD146-PE from Invitrogen as per manufacturer’s instructions. Markers expressions were characterized by acquiring samples in BD FACS-ARIA III (BD-Bioscience, Franklin Lakes, NJ, USA) using FACSDiva software. Compensation was performed by running either single color controls or BD CompBeads and analysis was performed using FlowJo software (BD-Bioscience, Franklin Lakes, NJ, USA).
3
Immunophenotyping of Mesenchymal Stem Cells
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On the 2nd passage, cells were analyzed by flow cytometry with BD FACSAria cell sorter (Becton Dickinson, Franklin Lakes, NJ, USA) using anti-mouse monoclonal antibodies: CD90 APC-Cy7 (BD Biosciences, cat. no. 561401, Franklin Lakes, NJ, USA), CD105 APC (Invitrogen, cat. no. 17-1051-82, Carlsbad, CA, USA), CD73 PE (BD Biosciences, cat. no. 550741, Franklin Lakes, NJ, USA), CD44 PE (BD Biosciences, cat. no. 553134, Franklin Lakes, NJ, USA), CD45 PE (Thermo Fisher Scientific, cat. no. MA1-10233, Waltham, MA, USA), CD34 Alexa Fluor® 647 (BD Biosciences, cat. no. 560230, Franklin Lakes, NJ, USA).
4
Characterization of UCSC Surface Markers
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UCSCs were obtained from Cyagen Biosciences Inc. (HUXUC-01001, Cyagen). The UCSCs were placed in a 6-well plate and cultured [31 (link)] in MEM (12571071, GIBCO) complete medium containing 10% fetal bovine serum (FBS; 10099141C, GIBCO) and 1% penicillin-streptomycin (15140122, GIBCO) with 5% CO2 at 37 °C. The media was changed every three days until the cells were confluent, and the cells were then digested and passaged using pancreatin (25200056, GIBCO).
Cells at passage 3 (P3) were collected to identify UCSCs surface markers including CD90-fitc (Invitrogen, Article No. 11-0909-41), CD29-pe-cy5 (BD PHOSFLOW, Article No. 559882), CD73-pe (Invitrogen, Article No. 12-0739-41), CD105-apc (Invitrogen, Article No. 17-1057-41), and HLA-apc-cy7 (Invitrogen, Article No. 47-9956-41). Cells were incubated in 100 μL staining solution at 4 °C for 30 min. We added 1 ml phosphate buffered saline (PBS) and centrifuged the cells at 400×g for 5 min. The supernatant was discarded, and the cells were suspended in 200 μL PBS for flow cytometry (FCM) analysis (Becton Dickinson, Aria II).
Cells at passage 3 (P3) were collected to identify UCSCs surface markers including CD90-fitc (Invitrogen, Article No. 11-0909-41), CD29-pe-cy5 (BD PHOSFLOW, Article No. 559882), CD73-pe (Invitrogen, Article No. 12-0739-41), CD105-apc (Invitrogen, Article No. 17-1057-41), and HLA-apc-cy7 (Invitrogen, Article No. 47-9956-41). Cells were incubated in 100 μL staining solution at 4 °C for 30 min. We added 1 ml phosphate buffered saline (PBS) and centrifuged the cells at 400×g for 5 min. The supernatant was discarded, and the cells were suspended in 200 μL PBS for flow cytometry (FCM) analysis (Becton Dickinson, Aria II).
5
Differentiation of ESCs to MSCs
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MSCs were derived from ESCs according to a protocol previously described (Zhang et al., 2015 (link)). Briefly, human ESC clones from embryoid bodies (EBs) were cultured in 90% α-MEM medium (Gibco) with 10% FBS (Ausbian, VS500T), 1% penicillin/streptomycin (Gibco), 5 ng/mL TGFβ (HumanZyme) and 10 ng/mL FGF2 (JPC) for 10 days until fibroblast-like cells emerged. The MSCs were sorted for CD73+/CD90+/CD105+ cells by FACS. Antibodies for sorting and characterization were CD73-PE (BD Biosciences, 550257), CD90-FITC (BD Biosciences, 555595), CD105-APC (eBioscience, 555748). IgG-FITC, IgG-PE, and IgG-APC were used as isotype controls. The differentiation abilities of MSCs to adipocytes, osteoblasts and chondrocytes were assessed by oil red O, Von Kossa and Toluidine blue staining, respectively (Duan et al., 2015 (link); Fang et al., 2018 (link); Yang et al., 2017 (link); Zhang et al., 2015 (link)).
6
Murine Mesenchymal Stem Cell Phenotyping
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For young and adult murine BD-MSCs, flow cytometry was performed using Negative markers: CD34, CD45, and positive markers: CD73, CD105, and CD90. Live cells were selected and the CD34-/CD45- population was further probed for CD73+/CD105+/90+. Antibodies used were CD34 FITC (Invitrogen), CD45 PerCP/Cy5.5 (Biolegend), CD73 PE (eBioscience) and CD105 APC (eBioscience) and CD 90 APC/Cy7 (Biolegend). Flow was run on LSRII and analyzed using FlowJo.
7
Flow Cytometric Characterization of BM-MSCs
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Flow cytometry was performed on BD-MSCs using markers for MSCs (Negative markers: CD34, CD45, and positive markers: CD73, and CD105) by using a BD LSR II Flow Cytometer . Live cells were selected and the CD34-/CD45- population, was gated for CD73+/CD105+. Antibodies used were CD34 FITC (Invitrogen), CD45 APC/cy7 (Biolegend), CD73 PE (eBioscience) and CD105 APC (eBioscience).
8
Phenotyping of Wharton's Jelly MSCs
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Surface antigen phenotyping of WJ-MSCs was assessed using flow cytometry (FACSCalibur™, BD Biosciences, USA) and analyzed with CellQuest Pro software. The cells were stained with anti-human antibodies labeled with phycoerythrin (PE), allophycocyanin (APC), fluorescein isothiocyanate (FITC), or PerCP. The specific antibodies used for staining included CD73-PE, CD44-FITC, CD29-PE, CD166-PE, CD45-FITC, CD34-PE, CD14-FITC, CD79a-APC, HLA-DR-PerCP (BD Pharmingen™, BD Biosciences, USA), CD105-APC (eBioscience™, ThermoFisher Scientific, USA), CD90-FITC, CD31-PE, HLA-ABC-APC, CD80-FITC, CD40-PE, and CD86-APC (Biolegend®, BioLegend Inc., USA). Before and following staining, samples were washed with 1× PBS. Isotype antibodies from the same manufacturers for mice or rats were used as controls.
9
Phenotypic Characterization of Stem Cells
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The stem cells were dissociated with trypsin, washed with cold PBS, and then separately stained with immunoglobulin G (IgG) or the following monoclonal antibodies conjugated to PE, PerCP, APC, or FITC: Epcam-PE, CD44-FITC, CD29-FITC, CD49f-FITC, CD73-FITC, CD105-APC, CD90-FITC, CD34-PerCP, CD31-FITC, CD45-FITC, and HLA-DR-FITC (all from eBioscience, USA). Upon being washed with PBS, the labeled cells were resuspended, and at least 105 events were acquired by using a BD Accuri™ C6 flow cytometer (BD, USA).
10
Multiparametric Phenotyping of MAPCs
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MAPCs were cultured, transferred to v bottom 96‐well plates and washed twice in flow cytometry staining buffer (PBS containing 2% FBS). Cell pellets were dissociated briefly by vortexing and antibodies for CD105 APC, ICAM1 PE, or PDL1 PE (eBioscience, Part of ThermoFisher Scientific) were added. Cells were incubated with surface antibodies for 30 minutes at 4°C, and then washed in flow cytometry staining buffer. Cells were then ready to acquire by flow cytometry on an Accuri C6. For intracellular staining, cells were incubated with 1X Brefeldin A (eBioscience, Part of ThermoFisher Scientific) for 4 hours before harvest. The intracellular FoxP3 kit was used per manufacturer's instructions (eBioscience, Part of ThermoFisher Scientific) to prepare cells for intracellular staining. Cells were blocked with 2% rat serum (eBioscience, Part of ThermoFisher Scientific) for 15 minutes to prevent nonspecific staining, and either IDO or COX‐2 antibodies (BD Biosciences, Berkshire, UK) were then added for 45 minutes. Cells were then washed in flow cytometry staining buffer and acquired using the Accuri C6 (BD Biosciences).
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