Clonally derived D1 mouse mesenchymal stromal cells (MSCs; American Type Cell Culture, CRL-12424) were used since they were used by a number of studies on cell-material interactions in the context of tissue regeneration [15 (link), 16 (link)]. Cells were cultured in complete high glucose Dulbecco’s Modified Eagle Media (DMEM; Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS; S11550, Atlanta Biologicals), 100 units/mL penicillin and 100 μg/mL streptomycin, and 2 mM GlutaMAX (Thermo Fisher Scientific). Cells were passaged when they reached 80% confluence in a 175 cm2 flask by detaching with trypsin-EDTA (Thermo Fisher Scientific). Passage numbers less than 15 were used for this study.
S11550
Manufactured by Atlanta Biologicals
Sourced in United States
The S11550 is a laboratory centrifuge that can be used to separate different components of a liquid sample. It operates at a maximum speed of 4,000 revolutions per minute and can accommodate various rotor types to handle different sample volumes and tube sizes.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
F0926, by BD (2 mentions)
B260300, by BD (2 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Htb 26, by American Type Culture Collection (1 mentions)
D6429, by Merck Group (1 mentions)
O1383, by Merck Group (1 mentions)
T75 culture flask, by Corning (1 mentions)
Mubmx 01001, by Cyagen (1 mentions)
Hela cell line, by American Type Culture Collection (1 mentions)
7 protocols using s11550
1
Clonal Mouse Mesenchymal Stromal Cells for Tissue Regeneration
2
Culture and Maintenance of TNBC Cell Lines
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The MDA-MB-231 and MDA-MB-157 human triple-negative breast cancer cell lines (HTB-26 and HTB-24, ATCC, Manassas, VA, USA). were grown in Dulbecco’s Modified Eagle Medium (DMEM; 11960-044, Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 2 mM glutamine (MIR6240, Mirus, Madison, WI, USA), 10% fetal bovine serum (FBS; S11550, Atlanta Biologicals, Nashville, TN, USA), 10,000 IU/mL penicillin and 10,000 µg/mL streptomycin (25300-054, Thermo Fisher Scientific) at 37 °C, 5% CO2 in a humidified incubator. Cell lines were previously tested for mycoplasma contamination. Experiments were performed in phenol red-free DMEM (31053-028, Thermo Fisher Scientific) supplemented with 2 mM glutamine and 10% FBS, except as noted.
3
Cultivation of Tick and Human Cell Lines
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I. scapularis embryonic cell lines ISE6 and IDE12 were cultured at 32°C with 1% CO2 in L15C-300 and L15C media, respectively. These growth media were supplemented with 10% heat-inactivated FBS (Sigma, F0926), 10% tryptose phosphate broth (TBP; BD, B260300), and 0.1% lipoprotein bovine cholesterol (LPBC; MP Biomedicals, 219147680)20 (link),95 (link).
HEK293 T cells were maintained in Dulbecco’s modified Eagle medium (DMEM; Sigma, D6429) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals; S11550) and 1x Glutamax. Cells were maintained in T75 culture flasks (Corning; 353136) at 33°C or 37°C in 5% CO2.
HEK293 T cells were maintained in Dulbecco’s modified Eagle medium (DMEM; Sigma, D6429) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals; S11550) and 1x Glutamax. Cells were maintained in T75 culture flasks (Corning; 353136) at 33°C or 37°C in 5% CO2.
4
Isolation and Culture of Syncytialized Placental Cells
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PHT cells were isolated from term placentas using the Kliman method
22 (link) involving DNAse/trypsin digestion and purification on a Percoll gradient as previously described.
12 (link) Cells were plated at a density of 2 x 106 per well for amino acid uptake assays or 2.75 × 106 cells per well in a 6‐well dish for siRNA gene silencing and subsequent experiments. Cells were cultured in equal volumes of Ham's F‐12 and high glucose DMEM, 10% fetal bovine serum (FBS; S11550, Atlanta Biologicals, Lawrenceville, GA), and antibiotics (penicillin, gentamicin and streptomycin) in 5% CO2, 95% atmospheric air at 37°C for 90 h. Culture media was changed daily.
Syncytialized PHT cells were treated with OA for 24 h starting at 66 h after plating. Culture media was changed to 1% FBS supplemented with bovine serum albumin (BSA, AK8909, Akron) in a 1:3 ratio with OA (100 μM, Sigma catalog # O1383). We have reported previously that this concentration of OA did not negatively affect PHT cell viability.
18 (link) PHT cells were studied at 90 h after plating.
22 (link) involving DNAse/trypsin digestion and purification on a Percoll gradient as previously described.
12 (link) Cells were plated at a density of 2 x 106 per well for amino acid uptake assays or 2.75 × 106 cells per well in a 6‐well dish for siRNA gene silencing and subsequent experiments. Cells were cultured in equal volumes of Ham's F‐12 and high glucose DMEM, 10% fetal bovine serum (FBS; S11550, Atlanta Biologicals, Lawrenceville, GA), and antibiotics (penicillin, gentamicin and streptomycin) in 5% CO2, 95% atmospheric air at 37°C for 90 h. Culture media was changed daily.
Syncytialized PHT cells were treated with OA for 24 h starting at 66 h after plating. Culture media was changed to 1% FBS supplemented with bovine serum albumin (BSA, AK8909, Akron) in a 1:3 ratio with OA (100 μM, Sigma catalog # O1383). We have reported previously that this concentration of OA did not negatively affect PHT cell viability.
18 (link) PHT cells were studied at 90 h after plating.
5
Culturing Tick and Human Cell Lines
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I. scapularis embryonic cell lines ISE6 and IDE12 were cultured at 32°C with 1% CO2 in L15C-300 and L15C media, respectively. These growth media were supplemented with 10% heat-inactivated FBS (Sigma, F0926), 10% tryptose phosphate broth (BD, B260300) and 0.1% lipoprotein bovine cholesterol (MP Biomedicals, 219147680) (20 (link), 105 (link)).
HEK293 T cells were maintained in Dulbecco’s modified Eagle medium (DMEM; Sigma, D6429) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals, S11550) and 1× Glutamax. Cells were maintained in T75 culture flasks (Corning, 353136) at 33°C or 37°C in 5% CO2.
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I. scapularis embryonic cell lines ISE6 and IDE12 were cultured at 32°C with 1% CO2 in L15C-300 and L15C media, respectively. These growth media were supplemented with 10% heat-inactivated FBS (Sigma, F0926), 10% tryptose phosphate broth (BD, B260300) and 0.1% lipoprotein bovine cholesterol (MP Biomedicals, 219147680) (20 (link), 105 (link)).
HEK293 T cells were maintained in Dulbecco’s modified Eagle medium (DMEM; Sigma, D6429) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals, S11550) and 1× Glutamax. Cells were maintained in T75 culture flasks (Corning, 353136) at 33°C or 37°C in 5% CO2.
6
Isolation and Characterization of Mouse and Human Bone Marrow MSCs
7
HeLa Cell Line Cultivation Protocol
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HeLa cell lines were purchased from ATCC. All cells were cultured at 37°C and 5.0% CO2 in Dulbecco’s modified Eagle medium (DMEM) (11995-065; Gibco) supplemented with 10% fetal bovine serum (FBS) (S11550; Atlanta Biologicals).
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