Lysolecithin (LPC)-mediated demyelination was performed as described in Kosaraju et al. (2020) (link). Briefly, 1-month-old WT mice were injected with LPC (Sigma, L1381) (1 μL of 1% solution in 1x PBS) using a stereotactic apparatus at two sites: +1.0 mm AP, +1.0 mm ML, −2.2 mm DV and +0.7 mm AP, +1.0ML, −2.2 DV. Animals were allowed to recover for 14 days.
L1381
Manufactured by Merck Group
Sourced in United States
The L1381 is a laboratory equipment product. It is used for performing various scientific experiments and analyses within a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
F8318, by Merck Group (1 mentions)
Recombinant mouse leptin, by Merck Group (1 mentions)
Af498, by R&D Systems (1 mentions)
Bovine serum albumin (bsa), by Corning (1 mentions)
Hydrophilic ptfe cell culture inserts, by Merck Group (1 mentions)
Milli q, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Corning (1 mentions)
7 protocols using l1381
1
Lysolecithin-Induced Demyelination Model
2
Lysopc-Sensitivity Screening of Lung Cells
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In order to screen lysoPC‐sensitive lung epithelial cells, the cell proliferation of large cell human lung carcinoma cells (NCI‐H460 and NCI‐H661) that expresses easily p53 mRNA detection, non‐SCLC cells (NCI‐H1299), lung ADC (A549 and NCI‐H1650), SCLC cells (NCI‐H1688), gene‐edited lung ADC epithelia (A549p53−), lung bronchial ADC epithelia (SPC‐A1 cells) and normal bronchial epithelia (HBE135‐E6E7) were measured using CCK8 (C0037, Beyotime, Shanghai, China) at 3, 6, 12, 24 and 48 h after treatment with exogenous lysoPC (L1381, Sigma‐Aldrich, MO, USA) at concentrations of 25, 50, 100 or 200 μM. Cells were cultured in full media consisting of RMPI 1640 (KGM31800‐500, KeyGEN, Jiangsu, China) containing 10% fetal bovine serum (F8318, Sigma‐Aldrich, MO, USA), penicillin 100 U/ml and streptomycin 100 mg/ml and in the cell incubator at 37°C with 5% CO2.
3
LPC-Induced Spinal Cord Injury Model
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Adult female mice at 7–8 weeks of age underwent laminectomy at Th11/Th12; 2 μl of 10 mg/ml lysophosphatidylcholine (LPC, L1381, Sigma Aldrich) dissolved in PBS was injected into the midline dorsal column at a depth of 0.5 mm13 (link). For administration of recombinant mouse leptin (Sigma Aldrich) or leptin neutralizing antibodies (AF498, R&D System), a cannula from Alzet osmotic pump (model No. 1002 or 1007D, Alzet Corp.) was placed at the thoracic spinal cord under the dura 3 days after LPC injection. The pump was filled with vehicle solution (PBS); recombinant mouse leptin (12 μg/kg body weight per day) or leptin neutralizing antibodies (10 μg/kg of body weight per day) were administered subcutaneously on the back.
4
LPC Spinal Cord Injury Modeling
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We described LPC injections more thoroughly elsewhere (10 (link)). Briefly, LPC (0.5-μl volume at 1%; L1381, Sigma-Aldrich) was injected into the T3-T4 ventral spinal cord white matter with a Hamilton 34-gauge needle using ketamine (200 mg/kg) and xylazine (10 mg/kg) to anesthetize mice. The spinal cord was injected lateral to the midline at 5° to a depth of 1.3 mm. Following needle withdrawal, the overlying musculature and skin were closed with sutures. Postoperative buprenorphine (0.05 mg/kg) was administered as an analgesic. After their recovery, mice were returned to their cages until appropriate time of euthanasia. Mice were euthanized via transcardial perfusion. Tissue was collected and fixed overnight with 4% paraformaldehyde (PFA) before cryoprotecting in a 30% sucrose solution, freezing, and cryosectioning onto slides.
5
Lysolecithin-Induced Demyelination in Mice
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Lysolecithin (LPC, L1381; Sigma, St. Louis, MO, USA) was dissolved in PBS at a final concentration of 1% (g/ml). P60 mice were anesthetized with Avertin by intraperitoneal injection and demyelination was induced by the focal injection of 2 μL 1% lysolecithin into the corpus callosum (1.00 mm posterior to bregma, 1.04 mm lateral to bregma, and 2.2 mm deep). LPC was delivered at a rate of 0.075 μL/min, and the needle remained in position for 10 min after LPC delivery and then was pulled out slowly. The wound was sutured, and the mice were placed on a heating pad until woken up.
6
Dorsal White Matter Demyelination Protocol
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7
Hydrogel-Based 3D Cell Culture Model
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All chemicals and consumables were used as received unless otherwise stated. Poly(ethylene glycol) diacrylate (PEGDA) (Mn = 700 g/mol), 2-hydroxy-2-methylpropiophenone (HMPP), phosphate buffered saline (PBS), bovine serum albumin (BSA), absolute ethanol (EtOH) and Corning Costar TC-treated multiple size 6 well plates were purchased from Sigma-Aldrich (St.
Louis, MO, USA). Two types of LPC were used for these studies: the "standard" LPC used for the ex vivo studies (Sigma, L1381) and the fluorescent analogue 1-(dipyrrometheneboron difluoride)undecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (TopFluor®Lyso PC) (Sigma, 810284P). ATTO 647-labeled maleimide was obtained from ATTO-TEC (Siegen, Germany).
Hydrophilic PTFE cell culture inserts with a diameter of 30 mm and a pore size of 0.4 µm were purchased from Merck Millipore (Darmstadt, Germany). Polystyrene templates with 0.5 mm, 1mm and 2 mm diameter cavities were self-made. All water used was highly purified and deionized using a Milli-Q ® ('ultrapure' water of "Type 1") integral water purification system from Merck Millipore (Darmstadt, Germany).
Louis, MO, USA). Two types of LPC were used for these studies: the "standard" LPC used for the ex vivo studies (Sigma, L1381) and the fluorescent analogue 1-(dipyrrometheneboron difluoride)undecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (TopFluor®Lyso PC) (Sigma, 810284P). ATTO 647-labeled maleimide was obtained from ATTO-TEC (Siegen, Germany).
Hydrophilic PTFE cell culture inserts with a diameter of 30 mm and a pore size of 0.4 µm were purchased from Merck Millipore (Darmstadt, Germany). Polystyrene templates with 0.5 mm, 1mm and 2 mm diameter cavities were self-made. All water used was highly purified and deionized using a Milli-Q ® ('ultrapure' water of "Type 1") integral water purification system from Merck Millipore (Darmstadt, Germany).
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