Medicon tissue disruptor

Manufactured by BD

Sourced in United States

The Medicon tissue disruptor is a laboratory instrument designed to mechanically disrupt and homogenize solid tissue samples. It utilizes high-speed rotational blades to efficiently break down and pulverize tissue samples, preparing them for further analysis or processing.

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Lab products found in correlation

3 protocols using medicon tissue disruptor

Isolation and Culture of Skin, Gut, and Immune Cells

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Whole-skin punch biopsies and cervix samples were incubated in 5U dispase (Life Technologies) overnight at 4°C followed by manual separation of epidermis and cervical epithelium from dermis or cervical submuocsa respectively followed by 90 min incubation in collagenase III (3 mg/ml; Worthington) with DNase (5 μg/ml; Roche) in RPMI 1640. Epidermal cell suspension was prepared by repeated pipetting. Dermis and submucosa were further processed by Medicon tissue disruptor (BD Biosciences) as previously described (Cheuk et al., 2014 (link)). Ileum biopsies were digested in collagenase II (0.25 mg/ml; Sigma-Aldrich) with DNase (0.2 mg/ml; Roche) in IMDM (Life Technologies) for 30–45 min. Complete RPMI medium was added and the cell suspension were subsequently passed through a 40 μm (gut) / 70 μm (skin or cervix) cell strainer (BD Bioscience). Peripheral blood mononuclear cells (PBMCs) were prepared by Ficoll (GE Healthcare) density separation.
P815 cells were purchased from ATCC and maintained in complete medium (RPMI 1640 supplemented with 10% fetal bovine serum [FBS], L-glutamine; all Hyclone). Recombinant IL-15, IL-1β, IL-2, IL-6, IL-7, IL-23, IL-12 and IFN-α (all R&D Systems) were stored at −80°C. Human collagen IV, phorbol 12-myristate 13-acetate (PMA), and ionomycin were purchased from Sigma.

Cheuk S., Schlums H., Gallais Sérézal I., Martini E., Chiang S.C., Marquardt N., Gibbs A., Detlofsson E., Introini A., Forkel M., Höög C., Tjernlund A., Michaëlsson J., Folkersen L., Mjösberg J., Blomqvist L., Ehrström M., Ståhle M., Bryceson Y.T, & Eidsmo L. (2017). CD49a Expression Defines Tissue-Resident CD8+ T Cells Poised for Cytotoxic Function in Human Skin. Immunity, 46(2), 287-300.

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Isolation of Skin T Cells from Different Compartments

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Peripheral mononuclear cells were isolated by Ficoll density gradient centrifugation (density 1.077; GE Healthcare Bio-Sciences, IL, USA). As for isolation of epidermal and dermal T cells, whole-skin specimens were incubated in 5 U/ml dispase (Life Technologies, CA, USA) overnight at 4 °C, and the epidermis was separated from the dermis. The epidermis was cut with scissors and incubated in collagenase III (3 mg/ml; Worthington Biochemical Corporation, NJ, USA) for 90 minutes with deoxyribonuclease (5 jig/ml; Sigma-Aldrich, MO, USA) in RPMI 1640 medium with 10% fetal bovine serum. A single-cell suspension was prepared by pipetting. The dermis was digested in the same way in collagenase III with deoxyribonuclease and further processed by a Medicon tissue disruptor (BD Biosciences, CA, USA). Short-term expansion culture was also applied to collect skin T cells from the whole-skin specimens or the epidermal/dermal specimens separated by dispase (Life Technologies) in the presence of 100 IU/ml of IL-2 (Wako, Osaka, Japan) and 20 ng/ml of IL-15 (Wako). The consistency of the T-cell phenotypes isolated from different body sites or different surgical techniques was also investigated in the subanalyses of our results (Supplementary Fig. 2).

Koguchi-Yoshioka H., Hoffer E., Cheuk S., Matsumura Y., Vo S., Kjellman P., Grema L., Ishitsuka Y., Nakamura Y., Okiyama N., Fujisawa Y., Fujimoto M., Eidsmo L., Clark R.A, & Watanabe R. (2021). Skin T cells maintain their diversity and functionality in the elderly. Communications Biology, 4, 13.

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Isolation and separation of skin cell types

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Fresh tissue samples were washed 2–3 times in PBS and incubated in 5 U/ml dispase (ThermoFisher Scientific) overnight at 4 °C. Epidermis was separated from dermis as described in ref. 41 (link). Epidermis was diced using scissors and then digested in Trypsin/EDTA Solution (ThermoFisher Scientific) for 15 minutes at 37 °C, from which CD45- (mainly composed of keratinocytes) and CD45 + cells (leukocytes) were separated using CD45 Microbeads with MACS MS magnetic columns (Milteney Biotec). Dermis was incubated in the enzyme mix from the whole skin dissociation kit (Milteney Biotec) for 3 hours according to manufacturer’s instructions and further processed by Medicon tissue disruptor (BD Biosciences). Dermal cell suspension was sorted sequentially through CD90 (fibroblasts), CD14 (macrophages) and CD3 (T cells) Microbeads with MACS MS magnetic columns according to manufacturer’s instructions (Milteney Biotec). And the CD90- CD14- CD3- dermal cells were also collected.

Li X., Li D., Wikstrom J.D., Pivarcsi A., Sonkoly E., Ståhle M, & Landén N.X. (2017). MicroRNA-132 promotes fibroblast migration via regulating RAS p21 protein activator 1 in skin wound healing. Scientific Reports, 7, 7797.

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