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Lamina propria dissociation kit

Manufactured by Miltenyi Biotec
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The Lamina Propria Dissociation Kit is a laboratory product designed to facilitate the isolation of cells from the lamina propria tissue. The kit contains the necessary reagents and tools to effectively dissociate this tissue and obtain a single-cell suspension for further analysis or experimentation.

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Lab products found in correlation

Gentlemacs dissociator, by Miltenyi Biotec (14 mentions) Gentlemacs octo dissociator, by Miltenyi Biotec (7 mentions) Percoll, by GE Healthcare (4 mentions) Facsdiva software, by BD (4 mentions) Celltrace violet, by Thermo Fisher Scientific (4 mentions) Facscanto 2, by BD (4 mentions) Facscalibur flow cytometer, by BD (3 mentions) Percoll gradient, by GE Healthcare (3 mentions) Lsrii flow cytometer, by BD (3 mentions) Percoll density gradient centrifugation, by GE Healthcare (2 mentions) Dithiothreitol (dtt), by Merck Group (2 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Anti f4 80, by Thermo Fisher Scientific (2 mentions) Lsr fortessa x 20 machine, by BD (2 mentions) Anti ly6g, by Miltenyi Biotec (2 mentions) Anti mouse cd16 cd32 antibody, by Thermo Fisher Scientific (2 mentions) Epcam, by Beckman Coulter (2 mentions) Epcam, by Thermo Fisher Scientific (2 mentions) Epcam, by BD (2 mentions)

Close spelling variants of this product

Mouse lamina propria dissociation kit (13 citations) Lamina propria kit (5 citations) Lamina propria dissociation kit for mice (2 citations) MACS lamina propria dissociation kit (2 citations) Miltenyi lamina propria dissociation kit (2 citations)

95 protocols using lamina propria dissociation kit

1

Non-classical Monocytes Homing in Colitis

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Colitis was induced in C57Bl/6J mice by supplementing their drinking water with 1 %–1.5 % dextran sulfate sodium (DSS, MP Biomedicals) for 7 days followed by 3 days of normal water.
Bone marrow cells were isolated from C57Bl/6J donor mice and cultured as mentioned above. After harvesting, NCMs were sorted and stained with CellTrace FarRed (Invitrogen) at 37°C for 20 min. One to 2.7 million labelled cells were injected into the ileocolic artery of mice with DSS colitis together with 100 µg anti‐β7 antibody (FIB504, BioXCell or Genentech), anti‐α4β7 antibody (DATK32, BioXCell) or IgG isotype (Sigma), respectively. After 2 h, lamina propria mononuclear cells (LPMCs) were isolated from the colon using the Lamina Propria Dissociation Kit (Miltenyi Biotec) according to the manufacturer's instructions followed by Percoll density gradient centrifugation (GE Healthcare). The LPMCs were stained for viable cells and/or Cd11b+ cells (Pacific Blue, M1/70, BioLegend), fixed with BD cell fix or FixPerm and the FarRed+ non‐classical monocytes homed to the gut were quantified by flow cytometry.
Sommer K., Heidbreder K., Kreiss L., Dedden M., Paap E., Wiendl M., Becker E., Atreya R., Müller T.M., Atreya I., Waldner M., Schürmann S., Friedrich O., Neurath M.F, & Zundler S. (2023). Anti‐β7 integrin treatment impedes the recruitment on non‐classical monocytes to the gut and delays macrophage‐mediated intestinal wound healing. Clinical and Translational Medicine, 13(4), e1233.
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2

Isolation of Intestinal Lamina Propria Mononuclear Cells

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To eliminate blood contamination of the small intestine, systemic perfusion with heparinized saline was performed prior to harvesting its tissue or washing with phosphate-buffered saline (PBS). The Lamina Propria Dissociation Kit (130-097-410; Miltenyi Biotec, Bergisch Gladbach, Germany) was used for the isolation of intestinal lamina propria (LPL) mononuclear cells according to the instructions. Cell pellets were resuspended in 40% Percoll® and slowly added to the top of a centrifuge tube containing 5 mL of 60% Percoll® at the bottom; washed twice with 2% FBS/PBS and centrifuged (420× g, 20 min) after density gradient to obtain LPL mononuclear cells.
Okamura T., Hamaguchi M., Mori J., Yamaguchi M., Mizushima K., Abe A., Ozeki M., Sasano R., Naito Y, & Fukui M. (2022). Partially Hydrolyzed Guar Gum Suppresses the Development of Sarcopenic Obesity. Nutrients, 14(6), 1157.
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3

Isolation of Lymphocytes from Spleen, mLN, and Intestine

Cited 1 time
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Single-cell suspensions of spleen and mesenteric LNs (mLNs) were prepared as previously described (Yeh et al., 2011 (link)) and resuspended in R10 medium (RPMI 1640 with 10% FCS, 2 mM l-glutamine, 100 IU/ml penicillin, 100 µg/ml streptomycin, 1× nonessential amino acids, 1 µM sodium pyruvate, and 50 µM β-mercaptoethanol). Large intestine lamina propria lymphocytes were isolated using the Lamina Propria Dissociation Kit (Miltenyi Biotec) according to the manufacturer’s instructions. Cells were then isolated by centrifugation over a Percoll gradient, washed, and resuspended in R10 media.
Shin B., Kress R.L., Kramer P.A., Darley-Usmar V.M., Bellis S.L, & Harrington L.E. (2018). Effector CD4 T cells with progenitor potential mediate chronic intestinal inflammation. The Journal of Experimental Medicine, 215(7), 1803-1812.
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4

Mouse Colon Lamina Propria Isolation

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Mouse colons were dissected and cut into 1 cm pieces within a petri dish. These colon segments were then submerged in an HBSS solution. To isolate lamina propria cells, we utilized the Lamina Propria Dissociation Kit (#130-097-410) from Miltenyi Biotec, following the manufacturer’s instructions.
Peng H.Y., Wang L., Das J.K., Kumar A., Ballard D.J., Ren Y., Xiong X., de Figueiredo P., Yang J.M, & Song J. (2023). Control of CD4+ T cells to restrain inflammatory diseases via eukaryotic elongation factor 2 kinase. Signal Transduction and Targeted Therapy, 8, 415.
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5

Isolation of Murine Lymphocytes

Cited 2 times
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Single-cell suspensions of spleen and lymph nodes were prepared as previously described 9 (link), 10 (link) and resuspended in R10 medium (RPMI 1640 with 10% FBS, 2 mM L-glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 10 mM HEPES, 1 mM sodium pyruvate, and 50 μM β-mercaptoethanol). Colon lamina propria lymphocytes were prepared using the Lamina Propria Dissociation Kit (Miltenyi Biotec, Auburn, CA) according to the manufacturer’s instructions. Cells were then isolated using a 40/75% discontinuous Percoll gradient, washed, and resuspended in R10 medium.
Yang W., Gibson S.A., Yan Z., Wei H., Tao J., Sha B., Qin H, & Benveniste E.N. (2020). Protein Kinase 2 (CK2) Controls CD4+ T-cell Effector Function in the Pathogenesis of Colitis. Mucosal immunology, 13(5), 788-798.
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6

Adoptive T Cell Transfer Colitis Model

Cited 1 time
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CD62LhiCD4+ T cells (106/ mouse) were intravenously injected to Rag1−/− or Rag1−/−Spp1−/− mice. Donor T cell apoptosis and population sizes were examined at day 3 and/or 6. The severity of colitis was evaluated by the ratio between weight and length of the colon, as well as histology staining with H&E and Alcian blue, at 4 weeks after T cell transfer. Histological scoring was performed as previously described46 (link). Colon infiltrated cells were isolated by using Lamina Propria Dissociation Kit (Miltenyi Biotec). To evaluate T cell proliferation, donor T cells were labeled with CellTrace Violet (Thermo Fisher Scientific inc.), and evaluated by flow cytometry 6 days after the injection. To examine an impact of OPN in maintaining B cell population, B cells (2× 106/ mouse) were adoptively transferred to Rag1−/− and analyzed the population size by flow cytometry at 4 weeks after the cell transfer.
Kanayama M., Xu S., Danzaki K., Gibson J.R., Inoue M., Gregory S.G, & Shinohara M.L. (2017). Skewing of the population balance of lymphoid and myeloid cells by secreted and intracellular osteopontin. Nature immunology, 18(9), 973-984.
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7

Intestinal Lamina Propria Isolation

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For consistency, the SI was measured and the most proximal third was harvested for flow cytometric analyses (regardless of differences in length). Fat and Peyer’s Patches were removed, and intestines were flushed with PBS and cut open longitudinally and into 1 cm segments. Mucus and epithelial cell removal were based on previously described methods [8 (link)]. Briefly, tissue segments were washed 20 min on a 200 rpm cell shaker in an HBSS solution of 5mM DTT (Sigma), followed by three 15 min washes in an HBSS solution of 5mM EDTA (Sigma). Segments were then digested with the mouse Lamina Propria Dissociation Kit used as directed (Miltenyi Biotec). After mechanical dissociation, samples were further mashed through a 70μM strainer.
Paustian A.M., Paez-Cortez J., Bryant S., Westmoreland S., Waegell W, & Kingsbury G. (2017). Continuous IL-23 stimulation drives ILC3 depletion in the upper GI tract and, in combination with TNFα, induces robust activation and a phenotypic switch of ILC3. PLoS ONE, 12(8), e0182841.
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8

Isolation and Purification of Immune Cells from Murine Spleen, Lymph Node, and Intestine

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Spleen (SP) and mesenteric lymph node (MLN) cells from each mouse in all groups were separated by grinding on filters. SP red blood cells were lysed using red blood cell lysis buffer (C3702; Beyotime, China). Single cell suspensions of SP and MLN were obtained through filters. Cells were washed twice with phosphate-buffered saline (PBS) (Zhongshanjinqiao, China) containing 2% fetal calf serum (FBS; as 2% FBS/PBS) (Gibco, Life Technologies, United States) through centrifugation.
Cell pellets were resuspended in the 2% FBS/PBS and were kept on ice for later use. Intestinal LPMCs were isolated in accordance with the Lamina Propria Dissociation Kit instructions (130-097-410; Miltenyi Biotec, Germany). Cell pellets were resuspended in 40% percoll (Ruitaibio, China) and added slowly to the upper part of centrifuge tubes, which had 5 mL of 80% percoll at the bottoms. LPMCs were obtained by washing twice with 2% FBS/PBS after density gradient centrifuging at 420 g for 20 min.
Guo H.X., Ye N., Yan P., Qiu M.Y., Zhang J., Shen Z.G., He H.Y., Tian Z.Q., Li H.L, & Li J.T. (2018). Sodium chloride exacerbates dextran sulfate sodium-induced colitis by tuning proinflammatory and antiinflammatory lamina propria mononuclear cells through p38/MAPK pathway in mice. World Journal of Gastroenterology, 24(16), 1779-1794.
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9

Isolation of Colonic Lamina Propria Leukocytes

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Colonic lamina propria leukocytes were isolated according to the Lamina Propria Dissociation Kit protocol from Miltenyi Biotec (Bergisch-Gladbach, Germany). Incubation was performed in a 37 °C incubator with sample agitation. Total leukocytes were purified from the digested tissue suspension using Percoll (GE Healthcare, Uppsala, Sweden) density (~1.041 g/mL) gradient centrifugation (20 min, 1800 rpm, room temperature, without rotor brake). Supernatant was discarded, the leukocyte pellet was washed once with phosphate buffered saline (PBS). Half the volume of the isolated leukocyte suspension was used for subsequent fluorescent activated cell sorting (FACS) analysis.
Gelmez E., Lehr K., Kershaw O., Frentzel S., Vilchez-Vargas R., Bank U., Link A., Schüler T., Jeron A, & Bruder D. (2022). Characterization of Maladaptive Processes in Acute, Chronic and Remission Phases of Experimental Colitis in C57BL/6 Mice. Biomedicines, 10(8), 1903.
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10

Isolation and Characterization of Intestinal Lamina Propria Immune Cells

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Lamina propria isolated from the small intestines by using the lamina propria dissociation kit (Miltenyi Biotec Inc) according to the manufacturer’s protocol. Cells from lamina propria were labeled with fluorochrome-conjugated antibodies, anti-F4/80 (eBioscience) and anti-Ly6G (Miltenyi Biotec). FACS was performed using a BD LSR Fortessa X-20 machine in the Janis V. Giorgi Flow Cytometry Core Facility at UCLA. For analysis and computational compensation of the data, BD FACS Diva software was used. Ten thousand events of live cells were gated.
Su F., GM A., Palgunachari M.N., White C.R., Stessman H., Wu Y., Vadgama J., Pietras R., Nguyen D., Reddy S.T, & Farias-Eisner R. (2020). Bovine HDL and Dual Domain HDL-Mimetic Peptides Inhibit Tumor Development in Mice. Journal of cancer research and therapeutic oncology, 8(1), 101.
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