The Effect of Age and Cell Culture Parameters on the Quantity and Function of Bone Marrow-derived Dendritic Cells
Abstract
Dendritic cells (DCs) are a group of bone marrow-derived cells that play a crucial role in innate and acquired immune responses. Bone marrow-derived dendritic cells (BMDC) are used in many studies, so the efficiency and purity of the differentiated cells are essential. This study aimed to investigate the effect of several parameters, including the age of mice, cell culture medium, and swirling of the culture plate, to increase the efficiency of the induced cells, considering the standard protocols.
Bone marrow-derived dendritic cells were induced from both juvenile and adult mice bone marrow cells. Then, the purity of CD11c+ cells was compared between juvenile mice BMDCs and adult mice BMDCs. Cells were cultured in an enriched and non-enriched medium, and some wells were swirled when changing the medium on the 3rd day. Then the effect of enriched medium and swirling before medium replacement were evaluated based on the expression of the CD11c marker.
The efficiency of DCs differentiation (CD11c+ cells) was higher when juvenile mouse bone marrow precursors were used compared to adult mice; using the enriched media with supplements and swirling the well before media replacement significantly affected the purity of immature CD11c+ cells.
Due to our results, using juvenile mice, an enriched culture medium, and physical removal of granulocyte cells could significantly improve the purity and efficiency of CD11c+ cells. Therefore, considering these three items in the production protocol of these cells can probably reduce the use of lymphocyte-removing antibodies and purification methods.
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8. Ganguly D, Haak S, Sisirak V, Reizis B. The role of dendritic cells in autoimmunity. Nat Rev Immunol. 2013;13(8):566-77
9. Adams AM, Chick RC, Vreeland TJ, Clifton GT, Hale DF, McCarthy PM, et al. Safety and efficacy of autologous tumor lysate particle-loaded dendritic cell vaccination in combination with systemic therapies in patients with recurrent and metastatic melanoma. Melanoma Res. 2021;31(4):378-88.
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12. Yazdani M, Gholizadeh Z, Nikpoor AR, Hatamipour M, Alani B, Nikzad H, et al. Vaccination with dendritic cells pulsed ex vivo with gp100 peptide-decorated liposomes enhances the efficacy of anti PD-1 therapy in a mouse model of melanoma. Vaccine. 2020;38(35):5665-77.
13. Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S, et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med. 1992;176(6):1693-702.
14. Lutz MB, Kukutsch N, Ogilvie AL, Rossner S, Koch F, Romani N, et al. An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods. 1999;223(1):77-92.
15. Roney K. Bone marrow-derived dendritic cells. Methods Mol. Biol. 2013;1031:71-6.
16. Liu J, Zhang X, Chen K, Cheng Y, Liu S, Xia M, et al. CCR7 Chemokine Receptor-Inducible lnc-Dpf3 Restrains Dendritic Cell Migration by Inhibiting HIF-1alpha-Mediated Glycolysis. Immunity. 2019;50(3):600-15 e15.
17. Posel C, Moller K, Frohlich W, Schulz I, Boltze J, Wagner DC. Density gradient centrifugation compromises bone marrow mononuclear cell yield. PloS One. 2012;7(12):e50293.
18. Rosca AM, Burlacu A. Isolation of a mouse bone marrow population enriched in stem and progenitor cells by centrifugation on a Percoll gradient. Biotechnol App Biochem. 2010;55(4):199-208.
19. Parish CR, Glidden MH, Quah BJ, Warren HS. Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation. Curr. Protoc. Immunol. 84:4.9.1 -4.9.13. 2009 Feb by John Wiley & Sons, Inc.;Chapter 4:Unit4 9.
20. Agrawal A, Gupta S. Impact of aging on dendritic cell functions in humans. Ageing Res Rev. 2011;10(3):336-45.
21. Chougnet CA, Thacker RI, Shehata HM, Hennies CM, Lehn MA, Lages CS, et al. Loss of Phagocytic and Antigen Cross-Presenting Capacity in Aging Dendritic Cells Is Associated with Mitochondrial Dysfunction. J Immunol. 2015;195(6):2624-32.
22. Grolleau-Julius A, Garg MR, Mo R, Stoolman LL, Yung RL. Effect of aging on bone marrow-derived murine CD
11c+CD4-CD8alpha- dendritic cell function. J Gerontol A Biol Sci Med Sci. 2006;61(10):1039-47.
23. Hoffman CM, Han J, Calvi LM. Impact of aging on bone, marrow and their interactions. Bone. 2019;119:1-7.
24. Zacca ER, Crespo MI, Acland RP, Roselli E, Nunez NG, Maccioni M, et al. Aging Impairs the Ability of Conventional Dendritic Cells to Cross-Prime CD8+ T Cells upon Stimulation with a TLR7 Ligand. PloS One. 2015;10(10):e0140672.
25. Willinghamm MD, Brodt MD, Lee KL, Stephens AL, Ye J, Silva MJ. Age-related changes in bone structure and strength in female and male BALB/c mice. Calcif. Tissue Int. 2010;86(6):470-83.
26. Liu L, Fan S, Lu Z, Chen Z, Chu C, Liu A, et al. An optimized method for the induction and purification of mouse bone marrow dendritic cells. J Immunol Methods. 2021:113073.
27. Dwivedi G, Chevrier A, Hoemann CD, Buschmann MD. Bone Marrow Progenitor Cells Isolated from Young Rabbit Trochlea Are More Numerous and Exhibit Greater Clonogenic, Chondrogenic, and Osteogenic Potential than Cells Isolated from Condyles. Cartilage. 2018;9(4):378-90.
28. Tong W, Tower RJ, Chen C, Wang L, Zhong L, Wei Y, et al. Periarticular Mesenchymal Progenitors Initiate and Contribute to Secondary Ossification Center Formation During Mouse Long Bone Development. Stem Cells. 2019 May;37(5):677-89. PubMed PMID: 30681752. Pubmed Central PMCID:
29. Maca RD. Inhibition of interleukin-2-induced proliferation of mouse T cells by normal mouse serum. J Biol Response Mod. 1983;2(4):360-8.
30. Martineau RS, Johnson JS. Normal mouse serum immunosuppressive activity: action on adherent cells. J Immunol. 1978;120(5):1550-3.
31. Mullbacher A, Woodhams CE, Tomaska LD, Pereira RA, Ashman RB, Blanden RV. Mouse serum as a medium supplement for murine immune responses in vitro. J Immunol Methods. 1985;76(1):17-26
32. Veit B, Michael JG. Characterization of an immunosuppressive factor present in mouse serum. J Immunol. 1973;111(2):341-51.
2. Segura E. Review of Mouse and Human Dendritic Cell Subsets. Methods Mol Biol. 2016;1423:3-15.
3. Palucka K, Banchereau J. Dendritic-cell-based therapeutic cancer vaccines. Immunity. 2013;39(1):38-48.
4. Santos PM, Butterfield LH. Dendritic Cell-Based Cancer Vaccines. J Immunol. 2018;200(2):443-9.
5. Bell GM, Anderson AE, Diboll J, Reece R, Eltherington O, Harry RA, et al. Autologous tolerogenic dendritic cells for rheumatoid and inflammatory arthritis. Ann Rheum Dis. 2017;76(1):227-34.
6. Nehmar R, Mariotte A, de Cauwer A, Sibilia J, Bahram S, Georgel P. Therapeutic Perspectives for Interferons and Plasmacytoid Dendritic Cells in Rheumatoid Arthritis. Trends Mol Med. 2018;24(4):338-47.
7. Spiering R, Jansen MAA, Wood MJ, Fath AA, Eltherington O, Anderson AE, et al. Targeting of tolerogenic dendritic cells to heat-shock proteins in inflammatory arthritis. J Transl Med. 2019;17(1):375.
8. Ganguly D, Haak S, Sisirak V, Reizis B. The role of dendritic cells in autoimmunity. Nat Rev Immunol. 2013;13(8):566-77
9. Adams AM, Chick RC, Vreeland TJ, Clifton GT, Hale DF, McCarthy PM, et al. Safety and efficacy of autologous tumor lysate particle-loaded dendritic cell vaccination in combination with systemic therapies in patients with recurrent and metastatic melanoma. Melanoma Res. 2021;31(4):378-88.
10. Bulgarelli J, Tazzari M, Granato AM, Ridolfi L, Maiocchi S, de Rosa F, et al. Dendritic Cell Vaccination in Metastatic Melanoma Turns "Non-T Cell Inflamed" Into "T-Cell Inflamed" Tumors. Front Immunol. 2019;10:2353.
11. Saberian C, Amaria RN, Najjar AM, Radvanyi LG, Haymaker CL, Forget MA, et al. Randomized phase II trial of lymphodepletion plus adoptive cell transfer of tumor-infiltrating lymphocytes, with or without dendritic cell vaccination, in patients with metastatic melanoma. J Immunother Cancer. 2021;9(5):12-8.
12. Yazdani M, Gholizadeh Z, Nikpoor AR, Hatamipour M, Alani B, Nikzad H, et al. Vaccination with dendritic cells pulsed ex vivo with gp100 peptide-decorated liposomes enhances the efficacy of anti PD-1 therapy in a mouse model of melanoma. Vaccine. 2020;38(35):5665-77.
13. Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S, et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med. 1992;176(6):1693-702.
14. Lutz MB, Kukutsch N, Ogilvie AL, Rossner S, Koch F, Romani N, et al. An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods. 1999;223(1):77-92.
15. Roney K. Bone marrow-derived dendritic cells. Methods Mol. Biol. 2013;1031:71-6.
16. Liu J, Zhang X, Chen K, Cheng Y, Liu S, Xia M, et al. CCR7 Chemokine Receptor-Inducible lnc-Dpf3 Restrains Dendritic Cell Migration by Inhibiting HIF-1alpha-Mediated Glycolysis. Immunity. 2019;50(3):600-15 e15.
17. Posel C, Moller K, Frohlich W, Schulz I, Boltze J, Wagner DC. Density gradient centrifugation compromises bone marrow mononuclear cell yield. PloS One. 2012;7(12):e50293.
18. Rosca AM, Burlacu A. Isolation of a mouse bone marrow population enriched in stem and progenitor cells by centrifugation on a Percoll gradient. Biotechnol App Biochem. 2010;55(4):199-208.
19. Parish CR, Glidden MH, Quah BJ, Warren HS. Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation. Curr. Protoc. Immunol. 84:4.9.1 -4.9.13. 2009 Feb by John Wiley & Sons, Inc.;Chapter 4:Unit4 9.
20. Agrawal A, Gupta S. Impact of aging on dendritic cell functions in humans. Ageing Res Rev. 2011;10(3):336-45.
21. Chougnet CA, Thacker RI, Shehata HM, Hennies CM, Lehn MA, Lages CS, et al. Loss of Phagocytic and Antigen Cross-Presenting Capacity in Aging Dendritic Cells Is Associated with Mitochondrial Dysfunction. J Immunol. 2015;195(6):2624-32.
22. Grolleau-Julius A, Garg MR, Mo R, Stoolman LL, Yung RL. Effect of aging on bone marrow-derived murine CD
11c+CD4-CD8alpha- dendritic cell function. J Gerontol A Biol Sci Med Sci. 2006;61(10):1039-47.
23. Hoffman CM, Han J, Calvi LM. Impact of aging on bone, marrow and their interactions. Bone. 2019;119:1-7.
24. Zacca ER, Crespo MI, Acland RP, Roselli E, Nunez NG, Maccioni M, et al. Aging Impairs the Ability of Conventional Dendritic Cells to Cross-Prime CD8+ T Cells upon Stimulation with a TLR7 Ligand. PloS One. 2015;10(10):e0140672.
25. Willinghamm MD, Brodt MD, Lee KL, Stephens AL, Ye J, Silva MJ. Age-related changes in bone structure and strength in female and male BALB/c mice. Calcif. Tissue Int. 2010;86(6):470-83.
26. Liu L, Fan S, Lu Z, Chen Z, Chu C, Liu A, et al. An optimized method for the induction and purification of mouse bone marrow dendritic cells. J Immunol Methods. 2021:113073.
27. Dwivedi G, Chevrier A, Hoemann CD, Buschmann MD. Bone Marrow Progenitor Cells Isolated from Young Rabbit Trochlea Are More Numerous and Exhibit Greater Clonogenic, Chondrogenic, and Osteogenic Potential than Cells Isolated from Condyles. Cartilage. 2018;9(4):378-90.
28. Tong W, Tower RJ, Chen C, Wang L, Zhong L, Wei Y, et al. Periarticular Mesenchymal Progenitors Initiate and Contribute to Secondary Ossification Center Formation During Mouse Long Bone Development. Stem Cells. 2019 May;37(5):677-89. PubMed PMID: 30681752. Pubmed Central PMCID:
29. Maca RD. Inhibition of interleukin-2-induced proliferation of mouse T cells by normal mouse serum. J Biol Response Mod. 1983;2(4):360-8.
30. Martineau RS, Johnson JS. Normal mouse serum immunosuppressive activity: action on adherent cells. J Immunol. 1978;120(5):1550-3.
31. Mullbacher A, Woodhams CE, Tomaska LD, Pereira RA, Ashman RB, Blanden RV. Mouse serum as a medium supplement for murine immune responses in vitro. J Immunol Methods. 1985;76(1):17-26
32. Veit B, Michael JG. Characterization of an immunosuppressive factor present in mouse serum. J Immunol. 1973;111(2):341-51.
| Files | ||
| Issue | Vol 21 No 3 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijaai.v21i3.9803 | |
| Keywords | ||
| Bone marrow cells Cell differentiation Dendritic cells Primary cell culture | ||
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How to Cite
1.
Golara M, Amiri MM, Moazzeni SM. The Effect of Age and Cell Culture Parameters on the Quantity and Function of Bone Marrow-derived Dendritic Cells. Iran J Allergy Asthma Immunol. 2022;21(3):300-312.
