Bone marrow-derived hMSCs were purchased from PromoCell GmbH (Germany). The multilineage differentiation capacity of the hMSCs was verified by PromoCell GmbH prior to purchase. hMSCs were seeded at 4000 cells per cm2 and cultured in mesenchymal stem cell growth medium (MSCGM) (PromoCell GmbH, Germany). hMSCs were incubated at 37 °C in a 5% CO2 atmosphere and the medium was changed every three days. The cells were harvested at approximately 80% confluency with 0.025% (w/v) trypsin-EDTA in PBS, centrifuged, and sub-cultured in MSCGM. Passage 6 hMSCs were used for all cell experiments.
Hmscs
Manufactured by PromoCell
Sourced in Germany
HMSCs are multipotent stem cells derived from human bone marrow. They have the ability to differentiate into various cell types, including osteoblasts, chondrocytes, and adipocytes.
Automatically generated - may contain errors
Lab products found in correlation
Mscgm, by PromoCell (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Mesenchymal stem cell growth medium 2, by PromoCell (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Mesenchymal stem cell growth medium, by PromoCell (2 mentions)
α mem, by Thermo Fisher Scientific (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Trypsin edta, by Merck Group (1 mentions)
Trypan blue, by Thermo Fisher Scientific (1 mentions)
Countess, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin antibiotic, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Mem neaa, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Hmscs, by American Type Culture Collection (1 mentions)
Hfob1, by American Type Culture Collection (1 mentions)
Zirconium oxychloride, by Merck Group (1 mentions)
Penstrep, by HiMedia (1 mentions)
Graphite powder, by Merck Group (1 mentions)
24 protocols using hmscs
1
Expansion and Characterization of hMSCs
2
Culturing Human Mesenchymal Stem Cells
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Bone marrow–derived hMSCs were purchased from PromoCell GmbH (Germany). The multilineage differentiation capacity of hMSCs was verified by PromoCell GmbH prior to purchase. hMSCs were seeded at 4000 cells per cm2 in T225 flasks and cultured in mesenchymal stem cell growth medium (MSCGM) (PromoCell GmbH, Germany). hMSCs were incubated at 37 °C in a 5% CO2 atmosphere and the medium was changed every three days. The cells were harvested at 80% confluency with 0.025% (w/v) trypsin–EDTA in PBS, centrifuged, and sub–cultured in MSCGM. Passage 6 hMSCs were used for all cell response experiments.
3
Expansion and Characterization of hMSCs
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Bone marrow-derived hMSCs were purchased from PromoCell GmbH (Germany). The multilineage differentiation capacity of the hMSCs was verified by PromoCell GmbH prior to purchase. hMSCs were seeded at 4000 cells per cm2 and cultured in mesenchymal stem cell growth medium (MSCGM) (PromoCell GmbH, Germany). hMSCs were incubated at 37 °C in a 5% CO2 atmosphere and the medium was changed every three days. The cells were harvested at approximately 80% confluency with 0.025% (w/v) trypsin-EDTA in PBS, centrifuged, and sub-cultured in MSCGM. Passage 6 hMSCs were used for all cell experiments.
4
Culturing Bone Marrow-Derived hMSCs
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Bone marrow-derived hMSCs were purchased from PromoCell GmbH (Germany). hMSCs were seeded at 4000 cells per cm2 in T225 flasks and cultured in mesenchymal stem cell growth medium (MSCGM) (PromoCell GmbH, Germany). hMSCs were incubated at 37 °C in a 5% CO2 atmosphere and the medium was changed every 3 d. The cells were harvested at 80% confluency with trypsin–EDTA (0.025% (w/v)) in PBS, centrifuged, and subcultured in MSCGM. Passage 6 hMSCs were used for all cell–material interaction experiments.
5
Culturing Bone Marrow-Derived hMSCs
6
Cell Recruitment in Scaffold-Hydrogel System
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A cell recruitment model was designed as described in our previous study[32 ]. In brief, the prepared scaffolds were placed in a donut-shaped alginate-gelatin hydrogel with human mesenchymal stem cells (hMSCs) derived from bone marrow (PromoCell, Germany). Then, the hMSCs were cultured in Mesenchymal Stem Cell Growth Medium 2 (PromoCell, Germany) for 7 and 14 days. After the scheduled culture period, the samples were stained with 4′,6-diamidino-2-phenylindole and observed using a confocal microscope.
7
hMSCs Culture on GelMA/Silica Scaffolds
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For cell culture, the GelMA/silanated silica scaffolds (5 mm3 × 5 mm3 × 2.4 mm3) were prepared and sterilized with 70% ethanol and UV light for 3 h each, and then placed in a culture medium overnight. Human mesenchymal stem cells derived from bone marrow (hMSCs-BM; Promo Cell, Heidelberg, Germany) were cultured in the composite scaffolds to observe the cellular behavior. hMSCs were cultured in mesenchymal stem cell growth medium 2 (Promo Cell, Heidelberg, Germany). Each scaffold is seeded with cells at a density of 5 × 104 and put in incubation (5% CO2 at 37°C), with medium exchange every second day. The cells were cultured in DMEM low glucose containing 50 μg mL−1 vitamin C and 10 mM β-glycerophosphate.
8
Expansion of Bone Marrow-Derived hMSCs
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Bone marrow-derived hMSCs were purchased from PromoCell GmbH (Heidelberg). The hMSCs were seeded at 4000 cells per cm2 in T225 flasks and cultured in mesenchymal stem cell growth medium (MSCGM) (PromoCell GmbH, Heidelberg). hMSCs were incubated at 37 °C and 5% CO2 and the medium changed every three days. The cells were harvested at 80% confluency with 0.025% (w/v) trypsin-EDTA (Sigma Aldrich, UK) in PBS, centrifuged and subcultured in MSCGM. Passage 6 hMSCs were used for all hydrogel encapsulation experiments.
9
Expansion and Attachment of Human Mesenchymal Stem Cells
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Human bone marrow mesenchymal stem cells
(hMSCs) (PromoCell GmbH, Germany) were expanded and maintained in
cell medium (Dulbecco’s modified Eagle’s medium (DMEM),
10% fetal bovine serum (FBS), 1% fungizone, 1% penicillin/streptomycin,
2 mMl -glutamine, and 0.5% nonessential amino acids (NEEA)
(100×)) at 37 °C with 5% CO2 for 5 days in a
75 cm2 Corning culture flask. When 80% cells were confluent,
they were trypsinized from the culture flask using trypsin–EDTA
spun down at 1000 rpm for 5 mins. Afterward, they were resuspended
in media before further processing. All reagents were acquired from
Sigma-Aldrich, U.K.
For cell attachment and morphology studies,
cells were seeded onto the cements at a density of 5000 cells/cm2 and incubated at 37 °C with 5% CO2 in cell
medium with no FBS for 4 h for cell attachment and morphology studies.
(hMSCs) (PromoCell GmbH, Germany) were expanded and maintained in
cell medium (Dulbecco’s modified Eagle’s medium (DMEM),
10% fetal bovine serum (FBS), 1% fungizone, 1% penicillin/streptomycin,
2 mM
(100×)) at 37 °C with 5% CO2 for 5 days in a
75 cm2 Corning culture flask. When 80% cells were confluent,
they were trypsinized from the culture flask using trypsin–EDTA
spun down at 1000 rpm for 5 mins. Afterward, they were resuspended
in media before further processing. All reagents were acquired from
Sigma-Aldrich, U.K.
For cell attachment and morphology studies,
cells were seeded onto the cements at a density of 5000 cells/cm2 and incubated at 37 °C with 5% CO2 in cell
medium with no FBS for 4 h for cell attachment and morphology studies.
10
Hypoxia-Induced Mesenchymal Stem Cell Proliferation
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Human umbilical cord derived mesenchymal stem cells (hMSCs; PromoCell) were grown in Dulbecco’s modified Eagle’s medium (DMEM; GIBCO) containing 10% fetal bovine serum (GIBCO) and 1% penicillin/streptomycin antibiotics (Invitrogen) at 37 °C in a 5% CO2 incubator with either 21% O2 (normoxia) or 1% O2 (hypoxia). The hypoxic condition culture cells were maintained in hypoxia after passage number 3. For cell number counting, cells were stained with Trypan blue (Invitrogen) and cell densities were determined with a Countess (Thermo Fisher Scientific) automated cell counter. The initial density of seeded cell 5000 cells per cm2. Cell number counting experiments were performed at 6 day after seeding for Fig. 1a and 3 day after seeding for Fig. 2a . Passage number 8 cells were used for metabolite treatment experiment.
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