Name: Economical and Efficient Protocol for Isolating and Culturing Bone Marrow-derived Dendritic Cells from Mice (Video) | JoVE Journal
Uploaded: 2022-07-01T12:30:00.000+00:00
Duration: 4 min 29 s
Description: Learn an economical protocol for isolating and culturing bone marrow-derived dendritic cells from mice using GM-CSF/IL-4. This method generates high-purity DCs for tumor immunity research in just seven days.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

This work was supported by Program of Tianjin Science and Technology Plan (20JCQNJC00550), Tianjin Health Science and Technology Project (TJWJ202021QN033 and TJWJ202021QN034).

All procedures were approved by the Nanjing Medical University Animal Care and Use Committee.
1. Isolation of bone marrow and preparation of BM cells
<ol>
	<li>Sacrifice C57BL/6 mice (18-22 g, 6-8 weeks old) via CO2 asphyxiation. Fix the mouse on the mouse operating table. Disinfect the surfaces with 70% ethanol.</li>
	<li>Cut the skin of the leg to expose the muscles and femoral artery. Clamp and tear off the femoral artery using two forceps, then pull the proximal end toward the abdomen. 
	NOTE: Do not cut the femoral artery directly. Otherwise, it will cause excessive bleeding and contaminate the field of vision.</li>
	<li>Cut all the muscles around the femur.</li>
	<li>Slowly stretch the lower limbs of the mouse outward until the sound of hip joint dislocation is heard and the femoral head prolapse is visible.</li>
	<li>Separate the lower limbs from the body using scissors along the inside of the femoral head.</li>
	<li>Cut the hind legs from the end of the knee joint to obtain a free and complete femur.</li>
	<li>Remove the muscles attached to the femur using gauze. 
	NOTE: Do not tear the muscles directly. The femur of mice is delicate, its integrity must be maintained.</li>
	<li>Immerse the femur in 75% alcohol for 2-5 min.</li>
</ol>
2. Induction culture of BMDC
<ol>
	<li>Rinse the residual alcohol with PBS. Use hemostatic forceps to clamp the middle and bottom part of the femur and another set to clamp the lower end of the femur. The hemostatic forceps are applied laterally to the femur, and the femur is separated from the epiphyseal line. 
	NOTE: The epiphyseal line is not easily visible until the femur fractures. This and the next steps are all performed in a sterile environment.</li>
	<li>Use a 1 mL syringe (needle: 0.6 mm x 25 mm) to penetrate the bone marrow cavity from the epiphyseal line break and rotate the needle to penetrate the femoral head and through the bone marrow cavity. Pulse and flush the bone marrow cavity using 1 mL of the complete medium containing GM-CSF/IL-4 until the bone becomes white. 
	NOTE: Complete culture medium: 10% FBS, 1% penicillin-streptomycin, 55 &#181;M &#946;-Mercaptoethanol, 10 ng/mL GM-CSF, and 10 ng/mL IL-4.</li>
	<li>Resuspend all the cells in 24 mL of complete culture medium (~5 x 105 cells/mL). After mixing, all the medium was seeded on a 6-well plate with 4 mL/well, then incubated at 37 &#176;C with 5% CO2 for 2 days. 
	NOTE: It is not necessary to lyse erythrocytes.</li>
	<li>Replace all the medium after 2 days with medium containing GM-CSF/IL-47. On the fourth, sixth, and eighth days, replace half of the medium with the complete medium containing 10 ng/mL GM-CSF/IL-4. 
	NOTE: In this step, suspended cells such as erythrocytes and lymphocytes are removed.</li>
	<li>Take pictures every day to record cell growth. Starting from day six, harvest one well of cells per day to count the cell number and detect CD11c, CD80, and MHC II expression6,7.</li>
</ol>
3. Flow cytometric detection of the expression of CD11c, CD80, and MHC II
<ol>
	<li>After washing the DCs with PBS, resuspend 1 x 106 cells in 100 &#181;L of FACS buffer, add 0.5 &#181;g of anti-mouse CD16/32 antibody, and incubate for 10 min on ice to block non-specific antigen sites.</li>
	<li>Add 1 &#181;g of Percp/cy5.5-CD11c, 0.5 &#181;g of PE-CD80, and 0.04 &#181;g of APC-MHC II antibodies, and incubate on ice for 30 min.</li>
	<li>Centrifuge at 1,000 x g for 3 min at 4 &#176;C, remove the supernatant, add 1 mL of 4% paraformaldehyde, fix for 30 min at 4 &#176;C, and re-suspend in 300 &#181;L of FACS buffer.</li>
	<li>Perform flow cytometry.</li>
</ol>

<ol><li>Huang, M. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antigen-loaded+monocyte+administration+induces+potent+therapeutic+antitumor+T+cell+responses%5BTitle%5D)+AND+%22Journal+of+Clinical+Investigation%22%5BJournal%5D)">Antigen-loaded monocyte administration induces potent therapeutic antitumor T cell responses.</a> Journal of Clinical Investigation. 130 (2), 774-788 (2020).</li>
<li>Wang, P., Dong, S., Zhao, P., He, X., Chen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Direct+loading+of+CTL+epitopes+onto+MHC+class+I+complexes+on+dendritic+cell+surface+in+vivo%5BTitle%5D)+AND+%22Biomaterials%22%5BJournal%5D)">Direct loading of CTL epitopes onto MHC class I complexes on dendritic cell surface in vivo.</a> Biomaterials. 182, 92-103 (2018).</li>
<li>Banchereau, J., Steinman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dendritic+cells+and+the+control+of+immunity%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Dendritic cells and the control of immunity.</a> Nature. 392 (6673), 245-252 (1998).</li>
<li>Jiang, P. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Galactosylated+liposome+as+a+dendritic+cell-targeted+mucosal+vaccine+for+inducing+protective+anti-tumor+immunity%5BTitle%5D)+AND+%22Acta+Biomaterialia%22%5BJournal%5D)">Galactosylated liposome as a dendritic cell-targeted mucosal vaccine for inducing protective anti-tumor immunity.</a> Acta Biomaterialia. 11, 356-367 (2015).</li>
<li>Shi, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Next-generation+immunotherapies+to+improve+anticancer+immunity%5BTitle%5D)+AND+%22Frontiers+in+Pharmacology%22%5BJournal%5D)">Next-generation immunotherapies to improve anticancer immunity.</a> Frontiers in Pharmacology. 11, 566401(2020).</li>
<li>Lutz, M. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+advanced+culture+method+for+generating+large+quantities+of+highly+pure+dendritic+cells+from+mouse+bone+marrow%5BTitle%5D)+AND+%22Journal+of+Immunological+Methods%22%5BJournal%5D)">An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow.</a> Journal of Immunological Methods. 223 (1), 77-92 (1999).</li>
<li>Son, Y. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+bulk-culture+method+for+generating+mature+dendritic+cells+from+mouse+bone+marrow+cells%5BTitle%5D)+AND+%22Journal+of+Immunological+Methods%22%5BJournal%5D)">A novel bulk-culture method for generating mature dendritic cells from mouse bone marrow cells.</a> Journal of Immunological Methods. 262 (1-2), 145-157 (2002).</li>
<li>Guo, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fusion+protein+vaccine+based+on+Ag85B+and+STEAP1+induces+a+protective+immune+response+against+prostate+cancer%5BTitle%5D)+AND+%22Vaccines%22%5BJournal%5D)">Fusion protein vaccine based on Ag85B and STEAP1 induces a protective immune response against prostate cancer.</a> Vaccines. 9 (7), 786(2021).</li>
<li>Olweus, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dendritic+cell+ontogeny%3A+A+human+dendritic+cell+lineage+of+myeloid+origin%5BTitle%5D)+AND+%22Proceedings+of+the+National+Academy+of+Sciences+of+the+United+States+of+America%22%5BJournal%5D)">Dendritic cell ontogeny: A human dendritic cell lineage of myeloid origin.</a> Proceedings of the National Academy of Sciences of the United States of America. 94 (23), 12551-12556 (1997).</li>
<li>Martin, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Concept+of+lymphoid+versus+myeloid+dendritic+cell+lineages+revisited%3A+both+CD8alpha%28-%29+and+CD8alpha%28%2B%29+dendritic+cells+are+generated+from+CD4%28low%29+lymphoid-committed+precursors%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Concept of lymphoid versus myeloid dendritic cell lineages revisited: both CD8alpha(-) and CD8alpha(+) dendritic cells are generated from CD4(low) lymphoid-committed precursors.</a> Blood. 96 (-), 2511-2519 (2000).</li>
<li>Anderson, D. A., Dutertre, C. A., Ginhoux, F., Murphy, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+models+of+human+and+mouse+dendritic+cell+development+and+function%5BTitle%5D)+AND+%22Nature+Reviews%3A+Immunology%22%5BJournal%5D)">Genetic models of human and mouse dendritic cell development and function.</a> Nature Reviews: Immunology. 21 (2), 101-115 (2021).</li>
<li>Vu Manh, T. P., Bertho, N., Hosmalin, A., Schwartz-Cornil, I., Dalod, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Investigating+evolutionary+conservation+of+dendritic+cell+subset+identity+and+functions%5BTitle%5D)+AND+%22Frontiers+in+Immunology%22%5BJournal%5D)">Investigating evolutionary conservation of dendritic cell subset identity and functions.</a> Frontiers in Immunology. 6, 260(2015).</li>
<li>Scheicher, C., Mehlig, M., Zecher, R., Reske, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dendritic+cells+from+mouse+bone+marrow%3A+in+vitro+differentiation+using+low+doses+of+recombinant+granulocyte-macrophage+colony-stimulating+factor%5BTitle%5D)+AND+%22Journal+of+Immunological+Methods%22%5BJournal%5D)">Dendritic cells from mouse bone marrow: in vitro differentiation using low doses of recombinant granulocyte-macrophage colony-stimulating factor.</a> Journal of Immunological Methods. 154 (2), 253-264 (1992).</li>
<li>Brasel, K., De Smedt, T., Smith, J. L., Maliszewski, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Generation+of+murine+dendritic+cells+from+flt3-ligand-supplemented+bone+marrow+cultures%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Generation of murine dendritic cells from flt3-ligand-supplemented bone marrow cultures.</a> Blood. 96 (9), 3029-3039 (2000).</li>
<li>Mayordomo, J. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((marrow-derived+dendritic+cells+pulsed+with+synthetic+tumour+peptides+elicit+protective+and+therapeutic+antitumour+immunity%5BTitle%5D)+AND+%22Nature+Medicine%22%5BJournal%5D)">marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.</a> Nature Medicine. 1 (12), 1297-1302 (1995).</li>
<li>Condon, C., Watkins, S. C., Celluzzi, C. M., Thompson, K., Falo, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((DNA-based+immunization+by+in+vivo+transfection+of+dendritic+cells%5BTitle%5D)+AND+%22Nature+Medicine%22%5BJournal%5D)">DNA-based immunization by in vivo transfection of dendritic cells.</a> Nature Medicine. 2 (10), 1122-1128 (1996).</li>
<li>Brunner, G. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Post-prandial+administration+of+the+insulin+analogue+insulin+aspart+in+patients+with+type+1+diabetes+mellitus%5BTitle%5D)+AND+%22Diabetic+Medicine%22%5BJournal%5D)">Post-prandial administration of the insulin analogue insulin aspart in patients with type 1 diabetes mellitus.</a> Diabetic Medicine. 17 (5), 371-375 (2000).</li>
<li>Koido, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Induction+of+antitumor+immunity+by+vaccination+of+dendritic+cells+transfected+with+MUC1+RNA%5BTitle%5D)+AND+%22Journal+of+Immunology%22%5BJournal%5D)">Induction of antitumor immunity by vaccination of dendritic cells transfected with MUC1 RNA.</a> Journal of Immunology. 165 (10), 5713-5719 (2000).</li>
<li>Jonasson, P. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Strength+of+the+porcine+proximal+femoral+epiphyseal+plate%3A+The+effect+of+different+loading+directions+and+the+role+of+the+perichondrial+fibrocartilaginous+complex+and+epiphyseal+tubercle+-+An+experimental+biomechanical+study%5BTitle%5D)+AND+%22Journal+of+Experimental+Orthopaedics%22%5BJournal%5D)">Strength of the porcine proximal femoral epiphyseal plate: The effect of different loading directions and the role of the perichondrial fibrocartilaginous complex and epiphyseal tubercle - An experimental biomechanical study.</a> Journal of Experimental Orthopaedics. 1 (1), 4(2014).</li>
<li>Labeur, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Generation+of+tumor+immunity+by+bone+marrow-derived+dendritic+cells+correlates+with+dendritic+cell+maturation+stage%5BTitle%5D)+AND+%22Journal+of+Immunology%22%5BJournal%5D)">Generation of tumor immunity by bone marrow-derived dendritic cells correlates with dendritic cell maturation stage.</a> Journal of Immunology. 162 (1), 168-175 (1999).</li>
<li>Hinkel, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Immunomodulatory+dendritic+cells+generated+from+nonfractionated+bulk+peripheral+blood+mononuclear+cell+cultures+induce+growth+of+cytotoxic+T+cells+against+renal+cell+carcinoma%5BTitle%5D)+AND+%22Journal+of+Immunotherapy%22%5BJournal%5D)">Immunomodulatory dendritic cells generated from nonfractionated bulk peripheral blood mononuclear cell cultures induce growth of cytotoxic T cells against renal cell carcinoma.</a> Journal of Immunotherapy. 23 (1), 83-93 (2000).</li>
</ol>

All authors declare they have no conflicts of interest and that they have nothing to disclose.

Humans and mice have different DC subsets, including classical DCs (cDCs, including cDC1s and cDC2s) plasmacytoid DCs (pDCs), and monocyte-derived DCs (MoDCs)9,10,11. It is generally accepted that cDC1s regulate cytotoxic T lymphocyte (CTL) responses to intracellular pathogens and cancer, and cDC2s regulate immune responses to extracellular pathogens, parasites, and allergens12. A significant number of DC...

Dendritic cells (DCs) are the most powerful antigen-presenting cells (APCs) for activating na&#239;ve T cells and inducing specific cytotoxic T lymphocyte (CTL) responses against infectious diseases, allergy diseases, and tumor cells1,2,3. DCs are the primary link between innate immunity and adaptive immunity and play an essential role in immunological defense and the maintenance of immune tolerance. In the last 40 years, many researchers have sought to define the subsets of DCs and their functions in inflammation and immunity. As per those studies, DCs develop along the myeloid and lymphoid lineages from bone marrow cells. Tumor vaccines have gained significant milestones in recent years and have a promising future. Mechanically, tumor vaccines modulate the immune response and prevent tumor growth by activating cytotoxic T lymphocytes using tumor antigens. The vaccine based on DCs plays an important role in tumor immunotherapy and has been identified as one of the most promising anti-tumor therapies1,4. In addition, DCs have been widely used in the testing of new molecular-targeted drugs and immune checkpoint inhibitors5.
Researchers urgently need a high number of high-purity DCs to further study the role of DCs. However, DCs are rare in various tissues and blood, accounting for only 1% of blood cells in humans and animals. In vitro culture of bone marrow dendritic cells (BMDC) is an important method for obtaining large amounts of DC cells. Meanwhile, The Lutz protocol for generating DCs from bone marrow has been widely used by researchers6. Although the protocol is effective in obtaining DC cells, it is complex and expensive, involving the addition of high concentrations of cytokines and the lysis of red blood cells.
In this study, we report a method for isolating almost all bone marrow cells from mouse bone marrow (BM) and inducing differentiation into BMDC after 7-9 days of incubation in vitro, with a lower concentration of GM-CSF and IL-4. This procedure only takes 10 min to harvest almost all bone marrow cells and to suspend them in a complete medium. In brief, we provide an efficient and cost-effective culturing method for BMDC in this research.

<table><tbody><tr><td>&amp;beta;-Mercaptoethanol</td><td>Solarbio</td><td>M8211</td><td></td></tr><tr><td>6-well plate</td><td>Corning</td><td>3516</td><td></td></tr><tr><td>APC-MHC II</td><td>Biolegend</td><td>116417</td><td></td></tr><tr><td>FBS</td><td>Gibco</td><td>10100</td><td></td></tr><tr><td>PE-CD80</td><td>Biolegend</td><td>104707</td><td></td></tr><tr><td>Penicillin-Streptomycin</td><td>Solarbio</td><td>P1400</td><td></td></tr><tr><td>Percp/cy5.5-CD11c</td><td>Biolegend</td><td>117327</td><td></td></tr><tr><td>PRMI-1640</td><td>Thermo</td><td>11875093</td><td></td></tr><tr><td>Recombinant Mouse GM-CSF</td><td>Solarbio</td><td>P00184</td><td></td></tr><tr><td>Recombinant Mouse IL-4</td><td>Solarbio</td><td>P00196</td><td></td></tr><tr><td>TruStain Fc PLUS (anti-mouse CD16/32) Antibody</td><td>Biolegend</td><td>156603</td><td></td></tr></tbody></table>

economical and efficient protocol for isolating and culturing bone marrow-derived dendritic cells from mice

Economical and Efficient Protocol for Isolating and Culturing Bone Marrow-derived Dendritic Cells from Mice

The demand for dendritic cells (DCs) is gradually increasing as immunology research advances. However, DCs are rare in all tissues. The traditional method for isolating DCs primarily involves inducing bone marrow (BM) differentiation into DCs by injecting large doses (&#62;10 ng/mL) of granulocyte-macrophage colony-stimulating factor/interleukin-4 (GM-CSF/IL-4), making the procedure complex and expensive. In this protocol, using all BM cells cultured in 10 ng/mL GM-CSF/IL-4 medium, after 3-4 half-culture exchanges, up to 2.7 x 107 CD11c+ cells&#160;(DCs) per mouse (two femurs) were harvested with a purity of 80%-95%. After 10 days in culture, the expression of CD11c, CD80, and MHC II increased, whereas the number of cells decreased. The number of cells peaked after 7 days of culture. Moreover, this method only took 10 min to harvest all bone marrow cells, and a high number of DCs were obtained after 1 week of culture.

The 1 x 107-1.7 x 107 cells were extracted from two femurs and were re-suspended in 24 mL of medium before being planted in a 6-well plate (Figure 1A). After 2 days, non-adherent cells were removed by completely changing the culture medium. Before changing the medium, a significant number of suspended cells were observed (Figure 1B). After 3 days of culture, small cell colonies began to form. On the sixth day, the size and number of colonie...

Here, we present an economical and efficient method to isolate and generate high-purity bone marrow-derived dendritic cells from mice after 7 days of culture with 10 ng/mL GM-CSF/IL-4.

The demand for dendritic cells (DCs) is gradually increasing as immunology research advances. However, DCs are rare in all tissues. The traditional method ...

economical-efficient-protocol-for-isolating-culturing-bone-marrow

Research

JoVE Journal

Immunology and Infection

This video demonstrates the procedure for differentiating myeloid dendritic cells from mouse bone marrow. Isolation of mouse tibia and femur, and processing of bone marrow are demonstrated. Pictures demonstrating cell morphology before and after differentiation, and figures depicting cell phenotype and IL-12 production following maturation using CpG are shown.

Culture of myeloid dendritic cells from bone marrow precursors

This video describes a protocol for the generation of dendritic cells--key antigen-presenting cells that activate T cells--from the bone marrow of an adult mouse.

Bone Marrow-Derived Dendritic Cells Generation: A Method to Generate Dendritic Cells from Mouse Bone Marrow

We describe a protocol for isolation and purification of neutrophils from mouse bone marrow by density gradient centrifugation and for neutrophil labeling using CellTracker dyes. This represents a simple, fast, reproducible and economical method for obtaining large numbers of neutrophils for downstream functional studies or adoptive transfer and tracking experiments.

Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments

Here we present a protocol that generates large amounts of murine monocytes from heterogeneous bone marrow for translational applications. In comparison to others, this new method helps reduce the number of sacrificed animals and lowers costs by avoiding expensive methods such as high gradient magnetic cell separation (MACS).

Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes

Distinct dendritic cell subsets exist as rare populations in lymphoid organs, and therefore are challenging to isolate in sufficient numbers and purity for immunological experiments. Here we describe a high efficiency, high yield method for isolation of all of the currently known major subsets of mouse splenic dendritic cells.

An Efficient and High Yield Method for Isolation of Mouse Dendritic Cell Subsets

Monocyte-derived DC (MoDC) can sense minor amounts of danger-associated molecules and are therefore easily primed. We provide a detailed protocol for the isolation of MoDC from blood and tumors and their activation with immune complexes while highlighting key precautions that should be considered in order to avoid their premature activation.

Isolation Protocol of Mouse Monocyte-derived Dendritic Cells and Their Subsequent In Vitro Activation with Tumor Immune Complexes

Here, we present a protocol to develop and characterize tolerogenic dendritic cells (TolDCs) and evaluate their immunotherapeutic utility.

Development and Functional Characterization of Murine Tolerogenic Dendritic Cells

The present protocol describes the isolation and culture of bone marrow-derived macrophages from mice.

Isolation and Culture of Bone Marrow-Derived Macrophages from Mice

Here, we describe in vitro culture conditions, isolation, and increased generation of extracellular vesicles (EVs) from Echinococcus granulosus. The small EVs were characterized by dynamic light scattering and transmission electron microscopy. The uptake by bone marrow-derived dendritic cells and their phenotypic modulation were studied using confocal microscopy and flow cytometry.

Isolation And Dendritic Cell-Uptake of Small Extracellular Vesicles from Echinococcus granulosus

We present our optimized high-throughput nucleofection protocol as an efficient way of transfecting primary human monocyte-derived dendritic cells with either plasmid DNA or siRNA without causing cell maturation. We further provide evidence for successful siRNA silencing of targeted gene RIG-I at both the mRNA and protein levels.

Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation

Learn an economical protocol for isolating and culturing bone marrow-derived dendritic cells from mice using GM-CSF/IL-4. This method generates high-purity DCs for tumor immunity research in just seven days.

Economical and Efficient Protocol for Isolating and Culturing Bone Marrow-derived Dendritic Cells from Mice

Explore More Videos