Mouse hematopoietic lineage depletion cocktail kit

Manufactured by STEMCELL

Sourced in Canada

The Mouse Hematopoietic Lineage Depletion Cocktail Kit is a laboratory reagent designed to remove specific cell types from mouse hematopoietic cell samples. The kit contains a mixture of antibodies that target lineage-specific surface markers, allowing for the depletion of differentiated hematopoietic cells, such as T cells, B cells, and myeloid cells, from the sample.

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Lab products found in correlation

Sca 1 magnetic beads, by Miltenyi Biotec (4 mentions) C57bl 6 mice, by Jackson ImmunoResearch (2 mentions) C57bl 6, by Jackson ImmunoResearch (1 mentions)

Close spelling variants of this product

Lineage depletion kit (2 citations)

4 protocols using mouse hematopoietic lineage depletion cocktail kit

Isolation of Mouse Cardiac Mesenchymal Stem Cells

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Mouse C-MSCs were isolated from the hearts of 2 to 3 months old C57BL/6
mice (The Jackson Laboratory, Bar Harbor, Maine) by a 2-step procedure as
previously described with modification (11 (link), 18 (link), 33 ). Briefly, in step 1, ventricular heart tissues
were minced to a size of 1 mm³ followed by digestion with 0.1%
collagenase IV and 1 U/mL Dispase in DMEM/F-12. The digested heart tissue was
seeded into 6 well plated coated with fibronectin/gelatin (0.5 mg fibronectin in
100 mL 0.1% gelatin). Cardiac explant cultures were maintained until small,
round, phase-blank cells migrated from adherent explants and proliferated on the
fibroblast layer. We then depleted hematopoietic cells using the mouse
hematopoietic lineage depletion cocktail kit (Stemcell Technologies) by magnetic
activated cell sorting (MACS) followed by enriching Sca-1 + cells with Sca-1
magnetic beads (Miltenyi Biotec Inc., Auburn, CA) as instructed by the
manufacturer’s protocol. The sorted Sca-1 cells were cultured in complete
medium (DMEM/F12 containing 10% fetal bovine serum (FBS), 200mmol/L L-Glutamine,
55nmol/L β-mercaptoethanol and 1% MEM non-essential amino acid).

Ju C., Shen Y., Ma G., Liu Y., Cai J., Kim I.M., Weintraub N.L., Liu N, & Tang Y. (2018). Transplantation of cardiac mesenchymal stem cell-derived exosomes promotes repair in ischemic myocardium. Journal of cardiovascular translational research, 11(5), 420-428.

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Isolation and Enrichment of Cardiac Mesenchymal Stem Cells

Cited 1 time

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C-MSC were isolated from the hearts of 2-month-old male C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME, USA) using a two-step protocol as described previously [22 (link),23 (link)]. Briefly, in step 1, ventricular tissue was minced into 1 mm³ size, digested in DMEM medium with 0.1% collagenase IV and 1 U/mL dispase for 20 min, and then cultured in fibronectin/gelatin (0.5 mg fibronectin in 100 mL of 0.1% gelatin) coated 6-well plates until the small round phase-bright cells migrated from the adherent explants and proliferated on the fibroblast layer. In step 2, Sca-1+ cells were enriched from the phase-bright cells by the mouse hematopoietic lineage-depletion cocktail kit (STEMCELL Technologies, Vancouver, Canada), followed by enrichment for Sca-1⁺ cells by magnetic-activated cell sorting (MACS) with Sca-1 magnetic beads (Miltenyi Biotec Inc., Auburn, CA, USA) according to the manufacturer’s protocols. Selected Sca-1 cells were cultured and maintained in complete DMEM medium containing 10% fetal bovine serum, 200 mmol/L L-glutamine, 55 nmol/L β-mercaptoethanol, and 1% MEM nonessential amino acids.

Zhang H., Shen Y., Kim I.M., Liu Y., Cai J., Berman A.E., Nilsson K.R., Weintraub N.L, & Tang Y. (2023). Electrical Stimulation Increases the Secretion of Cardioprotective Extracellular Vesicles from Cardiac Mesenchymal Stem Cells. Cells, 12(6), 875.

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Isolation and Enrichment of Mouse Cardiac Mesenchymal Stem Cells

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Mouse C-MSCs were isolated from 2 to 3 months old mouse hearts (C57BL/6, The Jackson Laboratory, Bar Harbor, Maine) by a 2-step procedure as previously described with modification [11 (link), 12 (link), 26 ]. Briefly, in step 1, ventricular heart tissues were minced into 1 mm³ size, and then digested with 0.1% collagenase IV and 1 U/mL Dispase in DMEM/F-12. The digested heart tissue was seeded into 6 well plate coated with fibronectin/gelatin (0.5 mg fibronectin in 100 mL 0.1% gelatin). After two weeks, the migrated round, phase-bright cells migrated from adherent explants were collected, and undergone hematopoietic cell depletion using the mouse hematopoietic lineage depletion cocktail kit (Stem Cell Technologies) by magnetic activated cell sorting (MACS) followed by cell enrichment with Sca-1 magnetic beads (Miltenyi Biotec Inc., Auburn, CA) according to the manufacturer's protocol. The sorted Sca-1 cells were cultured in complete medium (DMEM/F12 containing 10% fetal bovine serum (FBS), 200mmol/L L-Glutamine, 55nmol/L β-mercaptoethanol and 1% MEM non-essential amino acid).

Ju C., Li Y., Shen Y., Liu Y., Cai J., Liu N., Ma G, & Tang Y. (2018). Transplantation of cardiac mesenchymal stem cell derived exosomes for angiogenesis. Journal of cardiovascular translational research, 11(5), 429-437.

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Isolation and Characterization of Notch1-Deficient Cardiac Mesenchymal Stem Cells

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Mouse C-MSCs were obtained from Dr. Yaoliang Tang at Augusta University, which were isolated from the hearts of 2- to 3-month-old Notch1^flox mice (The Jackson Laboratory, stock number: 006951) according to the procedure as previously described (Ju et al., 2018 (link)). The isolated cells were purified using a mouse hematopoietic lineage depletion cocktail kit (Stemcell Technologies) and Sca-1 magnetic beads (MiltenyiBiotec Inc.) with magnetic activated cell sorting (MACS). These cells expressed the mesenchymal cell surface makers CD105, CD44, and CD140 by flow cytometric analyses (Ju et al., 2018 (link)). Cells were cultured in high-glucose DMEM medium supplemented with 10% fetal bovine serum (FBS), 200 mM L-glutamine, 55 nM β-mercaptoethanol, and 1% MEM non-essential amino acid. Before EV collection, culture medium was switched to medium supplemented with exosome-deleted FBS (Gibco) for 48 h. C-MSCs isolated from Notch1^flox mice were designated as C-MSCs^Notch1^FF.

Xuan W., Khan M, & Ashraf M. (2020). Extracellular Vesicles From Notch Activated Cardiac Mesenchymal Stem Cells Promote Myocyte Proliferation and Neovasculogenesis. Frontiers in Cell and Developmental Biology, 8, 11.

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