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Liberase blendzyme ci

Manufactured by Roche
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Liberase blendzyme CI is a laboratory product developed by Roche. It is a collagenase enzyme blend used for the dissociation and isolation of cells from various tissue sources.

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Lab products found in correlation

Medimachine, by BD (3 mentions) Dnase, by Merck Group (2 mentions) Ficoll gradient, by Merck Group (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Collagenase 4, by Merck Group (1 mentions) Percoll gradient, by GE Healthcare (1 mentions) Cellquest, by BD (1 mentions) Facscan, by BD (1 mentions) Uc10 4f10 11, by BD (1 mentions)

Spelling variants of this product

Liberase (1194 citations) Liberase Blendzyme (29 citations) Liberase CI (15 citations)

3 protocols using liberase blendzyme ci

1

Isolation of Cardiac Lymphocytes

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For isolating lymphocytes in the heart, we used the protocol described by Gutierrez and colleagues [34 (link)]. Briefly, hearts collected from 5 mice at day 20 dpi were minced, pooled, and incubated for 1h at 37°C with RPMI 1640, supplemented with NaHCO3, penicillin-streptomycin gentamicin, and 0.05 g/mL of liberase blendzyme CI (Roche, Basel, Switzerland). The tissues were processed in Medimachine (BD Biosciences) with phosphate buffered saline (PBS) containing 0.01% bovine serum albumin (BSA). After tissue digestion and washes the lymphocytes were isolated by Ficoll gradient (Sigma).
Pontes Ferreira C., Cariste L.M., Ferri Moraschi B., Ferrarini Zanetti B., Won Han S., Araki Ribeiro D., Vieira Machado A., Lannes-Vieira J., Gazzinelli R.T, & Vasconcelos J.R. (2019). CXCR3 chemokine receptor guides Trypanosoma cruzi-specific T-cells triggered by DNA/adenovirus ASP2 vaccine to heart tissue after challenge. PLoS Neglected Tropical Diseases, 13(7), e0007597.
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2

Isolating Liver and Heart Lymphocytes

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The perfused liver was lysed with collagenase buffer composed of 0.2 mg/mL collagenase IV (SIGMA), 0.02 mg/mL DNase (SIGMA), and 5% FBS. The leukocytes were separated on a 35% Percoll gradient (GE Healthcare), followed by centrifugation at 600 × g for 20 min and at 4°C. The pellet was suspended in RPMI 1640 (SIGMA) with 10% FBS (30 (link)). For the purification of the lymphocytes of the heart, we followed the protocol of Gutierrez et al. (31 (link)). Briefly, hearts collected from five mice at day 20 d.p.i. were minced, pooled, and incubated for 1 h at 37°C with RPMI 1640, supplemented with NaHCO3, penicillin–streptomycin gentamicin, and 0.05 g/mL of liberase blendzyme CI (Roche, Basel, Switzerland). The organs were processed in a Medimachine (BD Biosciences) in PBS containing 0.01% BSA. After tissue digestion and washes, cell viability was assessed by trypan blue exclusion, counted in a hemocytometer.
Ferreira C.P., Cariste L.M., Santos Virgílio F.D., Moraschi B.F., Monteiro C.B., Vieira Machado A.M., Gazzinelli R.T., Bruna-Romero O., Menin Ruiz P.L., Ribeiro D.A., Lannes-Vieira J., Lopes M.D., Rodrigues M.M, & de Vasconcelos J.R. (2017). LFA-1 Mediates Cytotoxicity and Tissue Migration of Specific CD8+ T Cells after Heterologous Prime-Boost Vaccination against Trypanosoma cruzi Infection. Frontiers in Immunology, 8, 1291.
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3

Characterization of Regulatory T Cells in Periapical Lesions

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Isolation and characterization of lesions’ Tregs were performed as described previously (22 (link)). Whole periapical tissues of molars apex were initially incubated in RPMI-1640 with 0·28 Wunsch units/ml of liberase blendzyme CI (Roche, Basel, Switzerland), and then processed with 0·05% DNase (Sigma-Aldrich, Steinhein, Germany) using Medimachine (BDbiosciences, San Diego, CA, USA); followed by cell viability analysis (Trypan blue) and cell count (Neubauer chamber). Cells were incubated with optimal dilution fluorochrome-conjugated antibodies against CD4 (FITC, clone GK1.5, dilution 1:200), FOXp3 (Alexa488, R16-715, 1:100), CCR4 (PE, 1G1, 1:100), CCR5, (PE, C34-3448, 1:100), CCR7, (PE, 4B12, 1:50), CCR8, (PE, 1055c, 1:50), CXCR3, (PE, 1C6/CXCR3, 1:100), RANKL, (PE, IK22-5, 1:100), IL-10, (PE, JES5-16E3, 1:100), TGF-b and (PE, TW7-16B4, :200), CTLA-4 (PE, UC10-4F10-11, 1:200) (BD Biosciences) and then analyzed by flow cytometry (FACScan and CellQuest; BD Biosciences). Results represent the number of cells ± SD in the periapical tissues of each mouse, normalized by the tissue weight, or the number of positive cells for each marker in CD4+FOXp3+ subpopulation.
Francisconi C.F., Vieira A.E., Biguetti C.C., Glowacki A.J., Favaro Trombone A.P., Letra A., Menezes Silva R., Sfeir C.S., Little S.R, & Garlet G.P. (2015). Characterization of the Protective Role of Regulatory T Cells in Experimental Periapical Lesions Development and Its Chemoattraction Manipulation as a Therapeutic Tool. Journal of endodontics, 42(1), 120-126.
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