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Ls006365

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LS006365 is a laboratory equipment product. It serves a core function related to laboratory operations, though a detailed description is not available while maintaining an unbiased and factual approach.

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Lab products found in correlation

Hank s balanced salt solution, by Thermo Fisher Scientific (3 mentions) Collagenase type a, by Merck Group (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) P0781, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) L glutamine, by Merck Group (1 mentions) D5671, by Merck Group (1 mentions) Ls005273, by Worthington (1 mentions) α1 antitrypsin, by Merck Group (1 mentions) Ultrapure bsa, by Thermo Fisher Scientific (1 mentions) Chromium next gem single cell 3 reagent kits v3, by 10x Genomics (1 mentions) Evos m7000 imaging system, by Thermo Fisher Scientific (1 mentions) Collagenase a, by Roche (1 mentions)

6 protocols using ls006365

1

Quantifying Cell-Specific Drug Delivery

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Apoe−/−(Cdh5-CreERT2;Apoe−/−mT/mG mice treated
with Tamoxifen) mice were injected with 1 mg/kg 7C1 formulated with
Alexa647-tagged siLuciferase (VasoRx, Inc.). Two hours after intravenous
injection with 1 mg/kg 7C1-siLuciferase Alexa647, the aorta, heart, and lungs
were dissected, rinsed in cold PBS, and sliced into small pieces. The finely
minced tissue was transferred to a digestion mix consisting of Hank’s
balanced salt solution (Gibco) + 1 mg/ml collagenase type A (Sigma 10103578001)
+ 0.5 mg/ml elastase (Worthington LS006365) for 3 hr at 37°C and
pipetting every 30 min. The cell suspension was passed through a 40 μm
filter. DAPI (Sigma D9542) was used to detect dead cells. A FACS machine (BD
FACSAria) was used to analyze GFP+7C1-siLuciferase
Alexa647+ cells.
Chen P.Y., Qin L., Li G., Wang Z., Dahlman J.E., Malagon-Lopez J., Gujja S., Cilfone N.A., Kauffman K.J., Sun L., Sun H., Zhang X., Aryal B., Canfran-Duque A., Liu R., Kusters P., Sehgal A., Jiao Y., Anderson D.G., Gulcher J., Fernandez-Hernando C., Lutgens E., Schwartz M.A., Pober J.S., Chittenden T.W., Tellides G, & Simons M. (2019). Endothelial TGF-β signalling drives vascular inflammation and atherosclerosis. Nature metabolism, 1(9), 912-926.
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2

Isolation of Mouse Aortic VSMCs

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VSMCs were isolated from mouse aorta by digesting with 1 mg/mL collagenase (Gibco, 17104-019) and 1 U/ml elastase (LS006365, Worthington) in serum-free Dulbecco’s Modified Eagle Medium (DMEM)(D5671, SIGMA), supplemented with 2 mM L-glutamine (G7513, SIGMA), 100U/mL penicillin and 100 µg streptomycin (P0781, SIGMA) for 10 min at 37 °C. Adventitia was removed and the remaining tissue digested in 2.5 mg/ml collagenase and 2.5 U/ml elastase for 2 h with regular trituration. Cells were further passaged and maintained in complete DMEM with 10% FCS at 37 °C and 5% CO2 in humidified conditions.
Aravani D., Foote K., Figg N., Finigan A., Uryga A., Clarke M, & Bennett M. (2020). Cytokine regulation of apoptosis-induced apoptosis and apoptosis-induced cell proliferation in vascular smooth muscle cells. Apoptosis, 25(9), 648-662.
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3

Quantifying Cell-Specific Drug Delivery

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Apoe−/−(Cdh5-CreERT2;Apoe−/−mT/mG mice treated
with Tamoxifen) mice were injected with 1 mg/kg 7C1 formulated with
Alexa647-tagged siLuciferase (VasoRx, Inc.). Two hours after intravenous
injection with 1 mg/kg 7C1-siLuciferase Alexa647, the aorta, heart, and lungs
were dissected, rinsed in cold PBS, and sliced into small pieces. The finely
minced tissue was transferred to a digestion mix consisting of Hank’s
balanced salt solution (Gibco) + 1 mg/ml collagenase type A (Sigma 10103578001)
+ 0.5 mg/ml elastase (Worthington LS006365) for 3 hr at 37°C and
pipetting every 30 min. The cell suspension was passed through a 40 μm
filter. DAPI (Sigma D9542) was used to detect dead cells. A FACS machine (BD
FACSAria) was used to analyze GFP+7C1-siLuciferase
Alexa647+ cells.
Chen P.Y., Qin L., Li G., Wang Z., Dahlman J.E., Malagon-Lopez J., Gujja S., Cilfone N.A., Kauffman K.J., Sun L., Sun H., Zhang X., Aryal B., Canfran-Duque A., Liu R., Kusters P., Sehgal A., Jiao Y., Anderson D.G., Gulcher J., Fernandez-Hernando C., Lutgens E., Schwartz M.A., Pober J.S., Chittenden T.W., Tellides G, & Simons M. (2019). Endothelial TGF-β signalling drives vascular inflammation and atherosclerosis. Nature metabolism, 1(9), 912-926.
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4

Enzymatic Modeling of Mitral Valve ECM

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Excised leaflets were enzymatically digested in order to model the disruption of valvular ECM components associated with MVP. Three solutions were considered: elastase only, collagenase only, and a mixture of elastase and collagenase. Porcine pancreatic elastase (Worthington Biochemical; LS006365) was dissolved into a phosphate-buffered saline (PBS) solution at a concentration of 1 unit/mL, and purified collagenase (Worthington Biochemical; LS005273) was dissolved into a PBS solution at a concentration of 64 units/mL [22 (link)]. The elastase-collagenase mixture consisted of 1 unit of elastase and 64 units of collagenase dissolved per mL of PBS. Leaflets were submerged in digestion solution and maintained at 37°C and 5% CO2 for up to 3 hours. Stereo microscope images were recorded every 15 min for the first hour and every 30 min for the final two hours. Custom image processing analyses were used to semi-automatically identify leaflet boundaries and areas over time. Immediately following enzymatic digestion, leaflets were washed in PBS and placed in a solution of α1-Antitrypsin (Sigma-Aldrich; A6150) at a concentration of 0.5 mg/ml for 45 minutes to arrest enzyme activity and prevent further digestion [23 (link), 24 (link)].
Bender J.M., Adams W.R., Mahadevan-Jansen A., Merryman W.D, & Bersi M.R. (2020). Radiofrequency ablation alters the microstructural organization of healthy and enzymatically digested porcine mitral valves. Experimental mechanics, 61(1), 235-251.
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5

Isolation of Aortic Smooth Muscle Cells

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The ascending aorta of Apoe−/− and TGFβR2iSMC-Apoe mice were dissected from the mice and rinsed in cold PBS. The tissue was opened longitudinally and sliced into small fragments roughly 2 mm in length. The finely minced tissue was transferred to a digestion mix consisting of Hank’s balanced salt solution (Gibco) + 1 mg/ml collagenase type A (Sigma 10103578001) + 0.5 mg/ml elastase (Worthington LS006365) for 3 hrs at 37°C and pipetting every 30 min. DAPI (10 μg/ml; Sigma D9542) was used to detect dead cells. The cell suspension was passed through a 40 μm filter before sorting. A FACS machine (BD FACSAria) was used to sort GFP+RFP− live cells. Single cells were sorted into 0.4% BSA/PBS.
Chen P.Y., Qin L., Li G., Malagon-Lopez J., Wang Z., Bergaya S., Gujja S., Caulk A.W., Murtada S.I., Zhang X., Zhuang Z.W., Rao D.A., Wang G., Tobiasova Z., Jiang B., Montgomery R.R., Sun L., Sun H., Fisher E.A., Gulcher J.R., Fernandez-Hernando C., Humphrey J.D., Tellides G., Chittenden T.W, & Simons M. (2020). Smooth muscle cell reprogramming in aortic aneurysms. Cell stem cell, 26(4), 542-557.e11.
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6

Single-cell transcriptomics of aortic cells

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Ascending thoracic aortas were harvested from euthanized P45 Fbn1mgR/mgR and WT mice (n = 3 per genotype), and losartan- and vehicle-treated Fbn1mgR/mgR mice (n = 4 and 3, respectively). Aortic tissues of each genotype and treatment arm were pooled together, cleaned, minced, and digested in Hank’s balanced salt solution (14170112, Gibco) containing elastase (0.5 mg/mL, LS006365, Worthington) and collagenase A (10103578001, Roche) for 1 hour at 37°C and then passed through a 40 μm filter. Single-cell suspensions were incubated with eBioscience 1× RBC Lysis Buffer (00-4333-57, Invitrogen) for 3 minutes and collected in PBS containing 1% UltraPure BSA (AM2616, Invitrogen). After confirming cell count and viability using an EVOS M7000 imaging system (Invitrogen), each sample was immediately processed by the Genomics Core at Icahn School of Medicine at Mount Sinai according to the manufacturer’s instructions (Chromium Next GEM Single Cell 3′ Reagent Kits v3.1, 10× Genomics).
Sun Y., Asano K., Sedes L., Cantalupo A., Hansen J., Iyengar R., Walsh M.J, & Ramirez F. (2023). Dissecting aortic aneurysm in Marfan syndrome is associated with losartan-sensitive transcriptomic modulation of aortic cells. JCI Insight, 8(10), e168793.
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