BMSCs (ScienCell Research Laboratories, San Diego, CA, USA) were cultured in mesenchymal stem cell medium (ScienCell Research Laboratories) and maintained in a 5% CO2 humidified incubator (Thermo Scientific, NC, USA) at 37°C. A549 cells (Cell Bank, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; Cat# TCHu150, China) were cultured in DMEM/F-12 medium (Gibco, 12500-062, USA) supplemented with 10% fetal bovine serum (Hyclone, USA), 100 units/mL penicillin, and 100 mg/mL streptomycin and maintained in a 5% CO2 humidified incubator (Thermo Scientific) at 37°C.
Bmscs
Manufactured by ScienCell
Sourced in United States
BMSCs are primary bone marrow-derived mesenchymal stem cells. They are multipotent progenitor cells capable of differentiating into various cell types, including osteoblasts, chondrocytes, and adipocytes.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Alpha medium, by Corning (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Mesenchymal stem cell medium, by ScienCell (1 mentions)
A549 cells, by Thermo Fisher Scientific (1 mentions)
Co2 humidified incubator, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Cs150fnx, by Hitachi (1 mentions)
Bmscs, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
Huvecs, by ScienCell (1 mentions)
Endothelial cell growth supplement (ecgs), by ScienCell (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Mscs medium, by ScienCell (1 mentions)
Human bmscs, by ScienCell (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by ScienCell (1 mentions)
Hnpcs, by ScienCell (1 mentions)
10 protocols using bmscs
1
Culturing BMSCs and A549 Cells
2
BMSC Exosome Isolation Protocol
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BMSCs were obtained from ScienCell Research Laboratories (CA, USA), and were cultured in MSC medium containing 10% exosome-depleted FBS (Thermo Fisher, cat. A25904DG, MA, USA). Conditioned medium of BMSCs was gathered and centrifuged (2500 × g for 30 min, 8500 × g for 30 min) at 4 °C to remove cell debris, followed by centrifugation at 120,000 × g for 140 min (4 °C) using a Hitachi ultracentrifuge CS150FNX (Tokyo, Japan). The pellets were then washed by phosphate-buffered saline (PBS) under centrifugation of 120,000 × g (4 °C) for 70 min and stored at −80 °C.
3
Isolation of Mitochondria from Human BMSCs
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Human bone marrow-derived stromal cells (BMSCs) were purchased from ScienCell Inc. (Cat #: 7500). Cells were resuspended in mitochondria buffer (70 mM sucrose, 1 mM EGTA, 210 mM sorbitol, 10 mM MOPS [pH 7.4])41 (link) and then, the supernatant was collected after centrifugation for 10min (450g, 4 °C). For total fraction, 54 ul of the supernatant was collected after centrifugation for 10 min (550 g, 4 °C). The rest of supernatant was further centrifuged at 9500g for 10min at 4 °C and then, the supernatant was collected for the cytosol fraction. For mitochondria fraction, the pellet was resuspended in mitochondrial buffer, incubated on ice for 15 min, and centrifuged at 500 g for 10 min at 4 °C. The supernatant was further centrifuged at 9500g for 10min at 4 °C. Finally, the pellet was collected and resuspended in 54 ul mitochondria buffer as mitochondria fraction.
4
Optimizing BMSC Response to Hypoxia
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Human bone marrow mesenchymal stem cells (BMSCs; ScienCell) were cultured in MEM (Minimum Essential Medium) Alpha Medium (Corning) and 10% fetal bovine serum (FBS, Gibco, NY, USA). BMSCs between passages 4 and 8 were used for the subsequent experiments. Human umbilical vein endothelial cells (HUVECs; ScienCell) were cultured in Endothelial Cell Medium (ECM; ScienCell) containing 5% FBS (ScienCell) and 1% endothelial cell growth supplement (ScienCell). For DFO stimulation, DFO was dissolved in distilled water, which was then diluted in culture medium before use. BMSCs at 80% confluency were incubated for 48 hours in complete medium with exosomes-free FBS and supplemented with 50 μM DFO (Sigma-Aldrich), 100 μM DFO, 200 μM DFO, 400 μM DFO or distilled water as a negative control.
5
Mesenchymal Stem Cell Differentiation under Tensile Stress
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BMSCs have been purchased from ScienCell research laboratories (San Diego, CA, USA, Cat #2320) and isolated from adult human bone marrow. ADSCs were isolated from adult human fat issues using the type I collagenase digestion method. We cultured MSCs using DMEM (Gibco, Grand Island, NY, USA) at 37 °C in a 5% CO2 atmosphere. 10% heat-inactivated fetal bovine serum (Gibco) and a 100 units/mL of penicillin (Gibco) and 100 μg/mL of streptomycin (Gibco) were added to the culture medium. The subsequent experiments used the fifth generation of cells.
After 48 h of cell passage, all inhibitors were applied for 24 h, followed by tensile stress. To maintain the working concentration during stretching, the medium including different kinds of inhibitors should be changed every 2 days.
Five main groups were established: undifferentiated MSCs grown in growth medium (GM group), MSCs applied by cyclic tensile stress for 7 days (CTS group), MSCs treated with RGDyk under cyclic tensile stress (RGDyk group), MSCs treated with cytochalasin D under cyclic tensile stress (CytoD group), and MSCs treated with verteporfin under cyclic tensile stress (VP group).
After 48 h of cell passage, all inhibitors were applied for 24 h, followed by tensile stress. To maintain the working concentration during stretching, the medium including different kinds of inhibitors should be changed every 2 days.
Five main groups were established: undifferentiated MSCs grown in growth medium (GM group), MSCs applied by cyclic tensile stress for 7 days (CTS group), MSCs treated with RGDyk under cyclic tensile stress (RGDyk group), MSCs treated with cytochalasin D under cyclic tensile stress (CytoD group), and MSCs treated with verteporfin under cyclic tensile stress (VP group).
6
Isolation and Expansion of BMSCs
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BMSCs were obtained from the scienCell, Beijing, China; cultivated in MEM medium containing l-glutamine and sodium bicarbonate and supplemented with 10% FBS and 1% penicillin/streptomycin, respectively. The cells were grown in 90 mm petrie dish in a regulated CO2 incubator (HMUSSE, SQ-80N, Shanghai, China) at 37 ℃. Cells were confluent to 80–90%, washed several times with PBS (pH 7.4), digested with 0.25% trypsin and used for counting, plate spreading.
7
Culture and Expansion of BMSCs and NIH3T3 Cells
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Human BMSCs were purchased from ScienCell Research Laboratories (ScienCell, Carlsbad, CA, USA) and cultured according to the manufacturer's instructions. Briefly, BMSCs were cultured in MSCs medium (ScienCell) containing 5% fetal bovine serum (FBS), 1% mesenchymal stem cell growth supplement, and 1% penicillin/streptomycin. BMSCs in passages three to six were used for the in vitro experiments. The NIH3T3 fibroblast cell line was obtained from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Beijing, China) and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco, Invitrogen, Carlsbad, CA, USA) with 10% FBS (Gibco) and 1% penicillin-streptomycin (Gibco). All cells were cultured at 37 °C with 5% CO2 concentration, and passaged every 2–3 days when they reached approximately 90% confluence.
8
Culturing HNPCs and bMSCs in DMEM
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9
Isolation and Characterization of Exosomes from Bone Marrow Mesenchymal Stem Cells
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Human bone marrow mesenchymal stem cells (BM-SCs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) and cultured in MSC medium supplemented with 10% exosome-depleted FBS (Thermo Fisher, cat. A25904DG, Waltham, MA, USA). Primary rat bone marrow mesenchymal stem cells were obtained from rat femur and cultured in DMEM medium supplemented with 10% exosome-depleted FBS. The differentiation potential of bone marrow mesenchymal stem cells were identified using alizarin red S staining and oil red O staining. After 72 h, the conditioned medium was collected and centrifuged (1000 × g for 10 min, 9000 × g for 30 min) at 4 • C to remove cell debris, followed by centrifugation at 3000 × g for 90 min (4 • C) after moving into the ultrafiltration centrifuge tube. Then Exosome Isolation Reagent (Invitrogen, Carlsbad, CA, USA) was used in accordance with the manufacturer's protocol to isolate exosomes. The exosome-enriched fraction was diluted with PBS and quantified using a Pierce BCA protein assay kit before further use. BMSCs used in our study is from 4-6 passages.
10
Culture and Characterization of Human BMSCs and HBMECs
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Human bone marrow mesenchymal stem cells (BMSCs; ScienCell) were cultured in MEM (Minimum Essential Medium) Alpha Medium (Corning) and 10% fetal bovine serum (FBS, Gibco, NY, USA). BMSCs between passage 4 were used for subsequent experiments. Human brain microvascular endothelial cells (HBMECs) were purchased from ScienCell Research Laboratories (ScienCell Research Laboratories, San Diego, CA, USA). Cells were cultured in endothelial cell medium (ScienCell Research Laboratories) and incubated in a humidi ed atmosphere at 5% CO2 and 37°C.
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