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Heat inactivated

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Heat-inactivated is a laboratory process that uses heat treatment to deactivate or kill microorganisms in a sample. This technique is commonly used to prepare samples for various analytical procedures, ensuring the safety and integrity of the testing environment.

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Lab products found in correlation

L glutamine, by Thermo Fisher Scientific (3 mentions) Vernier caliper, by Thomas Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Recombinant human il 4, by Thermo Fisher Scientific (2 mentions) Il 10, by Thermo Fisher Scientific (2 mentions) Ofanti cd40 ab clone 82111, by R&D Systems (2 mentions) Yssel s serum free t cell medium, by Gemini Bio (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Fetal calf serum (fcs), by Gemini Bio (1 mentions) Streptomycin sulfate, by Thermo Fisher Scientific (1 mentions) Bovine serum, by Gemini Bio (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) 24 well plate, by Corning (1 mentions)

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Heat-inactivated FBS (12 citations) Heat-inactivated fetal bovine serum (6 citations) AB-positive heat-inactivated human serum (5 citations) Heat-inactivated human AB serum (3 citations) Heat-inactivated horse serum (2 citations) Heat-inactivated adult bovine serum (2 citations) GemCell heat inactivated fetal bovine serum (2 citations)

6 protocols using heat inactivated

1

Leishmania major Infection Model

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The origin of the L. major Seidman (MHOM/SN/74/SD) (LmSd) and the L. major Friedlin V1 (MHOM/IL/80/Friedlin) (LmFn) strains has been previously described [5 (link), 38 (link)]. Promastigotes were grown at 26°C in medium 199 supplemented with 20% heat-inactivated FCS (Gemini Bio-Products), 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine, 40 mM Hepes, 0.1 mM adenine (in 50 mM Hepes), 5 mg/ml hemin (in 50% triethanolamine) and 1 mg/ml 6-biotin (M199/S). Infective-stage, metacyclic promastigotes were isolated from stationary cultures (5–6 days) by density gradient centrifugation, as described previously [39 (link)]. Mice were then inoculated with 1000 metacyclic promastigotes in the ear dermis by intradermal (i.d.) injection in a volume of 10 μl. Lesion development was monitored weekly by measuring the diameter of the ear nodule with a direct-reading Vernier caliper (Thomas Scientific) and pathology was scored using the following scale: 0=no ulcer, 1=ulcer, 2= ear half eroded, 3=ear completely eroded. For histology, ears were fixed at different times post-infection with 4% formaldehyde, cut in 5μm serial sections and stained with hematoxylin and eosin (H&E).
Charmoy M., Hurrell B.P., Romano A., Lee S.H., Ribeiro-Gomes F., Riteau N., Mayer-Barber K., Tacchini-Cottier F, & Sacks D.L. (2016). The Nlrp3 inflammasome, IL-1β, and neutrophil recruitment are required for susceptibility to a non-healing strain of Leishmania major in C57BL/6 mice. European journal of immunology, 46(4), 897-911.
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2

PBMC and B Cell Culture Optimization

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PBMCs were cultured at 2 × 105 cells/well in a volume of 200 μl in 96-well, round-bottom plates. B cells were cultured at 3 × 105 cells/well in a volume of 200 μl in 96-well, flat-bottom plates. Cells were cultured in Yssel’s Serum-Free T-Cell Medium (Gemini BioProducts) with 10% heat-inactivated FBS (Gemini Bio-Products), 2 mM L-glutamine (Life Technologies), and penicillin/streptomycin (Life Technologies). Cells were stimulated with combinations of recombinant human IL-4 (40 ng/ml) and IL-10 (50 ng/ml; PeproTech, Rocky Hill, NJ). For B cell cultures, 0.1 μg/ml ofanti- CD40 Ab (clone 82111; R&D Systems, Minneapolis, MN) was added.
Lin A.A., Freeman A.F, & Nutman T.B. (2018). IL-10 Indirectly Downregulates IL-4–Induced IgE Production by Human B Cells. ImmunoHorizons, 2(11), 398-406.
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3

Maintenance and Infection of L. major Seidman Strain

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The L. major Seidman strain (MHOM/SN/74/SD) (LmSd) was maintained as follows: promastigotes were grown at 26°C in medium 199 supplemented with 20% heatinactivated FCS (Gemini Bio-Products), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, 40 mM Hepes, 0.1 mM adenine (in 50 mM Hepes), 5 mg/mL hemin (in 50% triethanolamine), and 1 mg/mL 6-biotin (M199/S). Parasites expressing a green fluorescent protein (LmSd-GFP) were grown using the identical culture medium. Infective-stage, metacyclic promastigotes were isolated from stationary cultures (5–6 days) by density gradient centrifugation, as described previously55 (link). Mice were then inoculated with metacyclic promastigotes in the ear dermis by intradermal injection in a volume of 10 μL. Lesion development was monitored weekly by measuring the diameter of the ear nodule with a direct-reading Vernier caliper (Thomas Scientific). Lesion pathology was also evaluated and scored as follows: 0 = no ulceration, 1 = ulcer, 2 = half ear eroded, 3 = ear completely eroded.
Lee S.H., Kang B., Kamenyeva O., Ferreira T.R., Cho K., Khillan J.S., Kabat J., Kelsall B.L, & Sacks D.L. (2023). Dermis resident macrophages orchestrate localized ILC2-eosinophil circuitries to maintain their M2-like properties and promote non-healing cutaneous leishmaniasis. Research Square.
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4

Axenic Culture of E. histolytica Trophozoites

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E. histolytica strain HM1:IMSS trophozoites were grown axenically in TYI-S-33 (trypticase-yeast extract-iron-serum) medium supplemented with 1X Diamond's vitamins (SAFC Biosciences, Lenexa, KS, USA), 15% heat-inactivated bovine serum (Gemini Bio-Products, West Sacramento, CA), 100 U penicillin/ml and 100 mg streptomycin sulfate/ml (Gibco/Life Technologies, Grand Island, NY, USA), at 37°C in T-25 tissue culture flasks [31 (link)].
Linford A.S., Jiang N.M., Edwards T.E., Sherman N.E., Van Voorhis W.C., Stewart L.J., Myler P.J., Staker B.L, & Petri WA J.r. (2014). Crystal structure and putative substrate identification for the Entamoeba histolytica low molecular weight tyrosine phosphatase. Molecular and biochemical parasitology, 193(1), 33-44.
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5

PBMC and B Cell Culture Optimization

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PBMCs were cultured at 2 × 105 cells/well in a volume of 200 μl in 96-well, round-bottom plates. B cells were cultured at 3 × 105 cells/well in a volume of 200 μl in 96-well, flat-bottom plates. Cells were cultured in Yssel’s Serum-Free T-Cell Medium (Gemini BioProducts) with 10% heat-inactivated FBS (Gemini Bio-Products), 2 mM L-glutamine (Life Technologies), and penicillin/streptomycin (Life Technologies). Cells were stimulated with combinations of recombinant human IL-4 (40 ng/ml) and IL-10 (50 ng/ml; PeproTech, Rocky Hill, NJ). For B cell cultures, 0.1 μg/ml ofanti- CD40 Ab (clone 82111; R&D Systems, Minneapolis, MN) was added.
Lin A.A., Freeman A.F, & Nutman T.B. (2018). IL-10 Indirectly Downregulates IL-4–Induced IgE Production by Human B Cells. ImmunoHorizons, 2(11), 398-406.
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6

Immune Response to Parasite Exposure

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Upon blister formation, epidermal sheets were surgically removed, washed three times in sterile PBS and cultured in 24-well plates (Costar, Cambridge, MA, USA), one explant per well, in 05 mL complete media [RPMI 1640 (Gibco, Gaithersburg, MD, USA) supplemented with 10% heat-inactivated foetal calf serum (Gemini Bio-Products, West Sacramento, CA, USA), 200 mM L-glutamine, 100 IU/mL penicillin (Invitrogen) and 100 μg/mL streptomycin (Invitrogen)]. Explants were cultured in media alone or in the presence of five L3s per explant or 1000 mf per explant. The explants and crawl-out cells were harvested at 24, 48, 72, 96 h or 5 days and processed for mRNA assessment (five explants in media alone and five explants exposed to parasite, one to be harvested per day at each time point). Supernatants were also collected and assessed for cytokine production at each time point.
COTTON R.N., MCDONALD-FLEMING R., BOYD A., SPATES K., NUTMAN T.B, & SEMNANI R.T. (2015). Brugia malayi infective larvae fail to activate Langerhans cells and dermal dendritic cells in human skin. Parasite immunology, 37(2), 79-91.
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