Myenteric neurons were dissociated from mouse small intestine, using a previously described method19 (link)39 (link). Cell cultures were prepared by adding 50 μL of a constantly mixed cell suspension into 8-well chambers (30108, SPL, Saveen Werner, SE) prefilled with 450 μL of Neurobasal A (NBA) cell culture medium (NBA, Thermo Fisher Scientific, SE with 10%v/v fetal bovine serum FBS, BioWest, FR, 0.5 mM L-glutamine K0282 and 50 U/mL penicillin, 50 μg/mL streptomycin A2213, BioChrom AG). From each animal two 8-well chambers (69 mm2/well) were prepared. Fresh medium containing applicable experimental test agents was added to cultures after a 4 day equilibration culture period. Control wells were cultured in parallel. Cells were fixed in Stefanini’s fixative, rinsed in Tyrode solution, frozen, thawed and subjected to immunocytochemistry.
Streptomycin a2213
Manufactured by Harvard Bioscience
Streptomycin A2213 is a laboratory reagent used for the detection and quantification of streptomycin, an antibiotic compound. It is a standardized solution that can be used in various analytical techniques to measure the concentration of streptomycin in samples.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Harvard Bioscience (2 mentions)
L glutamine k0282, by Harvard Bioscience (2 mentions)
Neurobasal a, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
β mercaptoethanol m6250, by Merck Group (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
M csf, by R&D Systems (1 mentions)
2 protocols using streptomycin a2213
1
Isolation and culture of mouse myenteric neurons
2
Isolation and Differentiation of Human PBMCs
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Human peripheral mononuclear cells (PBMCs) were isolated from buffy coats (DRK-Blutspendedienst Hessen GmbH, 506838) by passage over a Leukocyte Separation Medium (PAA, J15-004) gradient. The donors were healthy German adults without known exposure to Leishmania parasites. After separation, PBMCs were washed and resuspended in complete medium (CM), consisting out of RPMI 1640 (R0883) supplemented with 10% heat inactivated fetal calf serum (F7524), 50 μM β-mercaptoethanol (M6250) (all from Sigma Aldrich), 2 mM L -glutamine (K 0282), 100 U/ml penicillin and 100 μg/ml streptomycin (A 2213), 10 mM HEPES (L 1613) (all from Biochrom). Monocytes were isolated by exploiting their ability to adhere to plastic, or by CD14 positive selection (Miltenyi, 130-050-201). Nonadherent or CD14-negative cells were collected and frozen in CM containing 40% fetal calf serum and 10% DMSO for lymphocyte proliferation assays. Monocytes were differentiated into human monocyte derived macrophages by addition of 10 ng/ml recombinant human macrophages colony stimulating factor (M-CSF) (R&D, 216-MC), for a period of 5 to 7 d at 37°C, 5% CO2.
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