Clodronate Liposomes utilize the endocytosis mechanism of macrophages to carry membrane-impermeable clodronate (clodronate) into the cell. In the presence of macrophage lysosomal phosphatase, clodronate dissolved in the aqueous phase of liposomes is released and accumulates inside the cell. When it reaches a certain concentration it can induce the macrophage to enter the apoptotic process, thus functioning as a macrophage scavenger.

Applications

Macrophages depletion

Clodronate Liposomes are suitable for a variety of injection methods, such as intravenous, intraperitoneal, subcutaneous, intranasal and testicular. The injection volume is related to the body weight of the mice, the injection cycle, the injection method and the purpose of the experiment.

Specifications

Components

Shipping and Storage

The product is stored at 4℃. Transported in ice packs. Cannot be frozen! Shelf life 6 months.

FAQ

Q: For liver macrophage clearance, perfusion is required. Do you have any suggestions regarding flow cytometry?

A: Without perfusion, flow cytometry is meaningless because the macrophages haven't been separated. How can we observe the differences? Flow cytometry suggests detecting F4/80 and CD11b. It is recommended to inject via the tail vein, and then take the liver 24 hours later for immunohistochemistry. Immunohistochemistry does not require perfusion.

Q: The control group of the intracellular macrophage clearance agent, the blank lipid particles, is this a single-vesicle lipid particle or a multi-vesicle lipid particle?

A: Multi-room type.

Q: What is the concentration of the macrophage depleting agent?

A: 5 milligrams per milliliter.

Q: What is the particle size of the liposomes in the Clodronate Liposomes (from Vrije Universiteit Amsterdam) product, which is an intracellular macrophage clearance agent?

A: 150nm to 3um.

Q: How should the Clodronate liposomes be stored and handled after receiving them?

A: After receiving the chlorophosphate liposomes, if they cannot be used immediately, they should be stored in a refrigerator at 4℃. Do not freeze! Use them as the original solution and do not dilute. The liposomes are prone to precipitation. It is recommended to gently mix them before use. Also, the chlorophosphate liposomes at 4℃ cannot be used immediately; they need to be returned to room temperature before injection.

Q: Can Clodronate liposomes be used for the removal of macrophages in vitro?

A: The product can be used for the removal of macrophages in vitro, but this method is more suitable for in vivo experiments. This is mainly because during in vitro culture, the chlorine phosphate released from dead cells or the chlorine phosphate "leaking" from liposomes will remain in the culture medium continuously. However, in vivo, the half-life of chlorine phosphate is very short and it will be quickly cleared by the kidneys. Although free chlorine phosphate will not enter the cells or liposomes, once they accumulate in the culture medium, they will gradually enter the cells.

Q: After intravenous injection of Clodronate liposomes, the animals died very quickly. What was the reason?

A: On one hand, it is possible that the animals were injected with an uneven lipid suspension. It is recommended to gently shake and mix it before use. Liposomes will settle over time outside the body. If multiple animals need to be injected with one dose over a long period of time, the liposomes will precipitate in the syringe, forming an uneven suspension. As a result, the dosage injected into the first animal will be significantly different from that injected into the last animal. On the other hand, it is also possible that the liposomes have not yet returned to room temperature after being taken out of the refrigerator.

Q: For the removal of macrophages in different organs or tissues, where should the injection be performed, what should be the injection dose, and how should the injection time and cycle be determined?

A: This involves the design of a specific experimental plan. We suggest that the client design the experimental plan themselves. We can provide them with some reference materials or help them find relevant literature that aligns with their experimental goals.

Q: How many days after the injection of Clodronate liposomes did the animals die?

A: This might be due to bacterial contamination during the operation. After the macrophages in the body are removed, the chances of infection with viruses, bacteria or yeast increase.

Q: After intravenous injection of Clodronate liposomes, the ED1+ cells in the spleen and liver of the rats did not completely disappear?

A: In rats, mature macrophages show double positivity for ED1 and ED2. However, there are some precursor cells without or with weak phagocytic function that are also ED1+, but present as ED2-. Therefore, all cells positive for ED2 can be completely eliminated, but cells positive for ED1 can only be partially eliminated. The rate of elimination depends on the ratio of precursor/mature macrophages.

Q: The clodronate liposomes failed to achieve the expected results?

A: The chlorophosphate liposomes are produced in large quantities. For each batch, the concentration of chlorophosphate and some possible contaminants are tested to ensure accuracy before they are sold to customers. Additionally, liposomes are susceptible to extreme temperatures. The correct transportation and storage method is at 4 to 8 degrees Celsius. The suspension should neither be frozen nor heated above 30 degrees Celsius. Once the goods are received, they need to be used within 3 months; otherwise, the usage effect may be affected.

Q: During intravenous injection, can the volume of the injection be increased?

A: When performing intravenous injection, the dosage should be based on the animal's weight and cannot exceed 0.1 mL per 10 mg. However, for intraperitoneal injection, the volume of the injection can be appropriately increased. For subcutaneous injection, the dosage should be determined based on the volume of the injection site.

Q: The macrophage deactivator product got frozen during transportation. Can it still be used?

A: It cannot be used.

Q: Are macrophage degraders anions?

A: Yes, an anion.

Q: Regarding the macrophage clearance agent, does the 5 mg/mL mentioned in the instructions refer to the concentration of the drug?

A: Yes, it refers to the concentration of the drug.

Documents:

Safety Data Sheet

40337_MSDS_HB250103_EN.pdf

Manuals

40337-Manual-Ver.EN20260128

Related Blog

Clodronate Liposomes | Lesson 1: Injection Methods and Administration Strategies

Clodronate Liposomes | Lesson 2: Liver

Clodronate Liposomes | Lesson 3: Lung Macrophages

Clodronate Liposomes | Lesson 4: Intestine

Clodronate Liposomes | Lesson 5: Brain

Clodronate Liposomes | Lesson 6: Applications in C57BL/6 Mice

Clodronate Liposomes | Lesson 7: Peripheral Blood

Citations & References:

[1] Xiong X, Chen S, Shen J, et al. Cannabis suppresses antitumor immunity by inhibiting JAK/STAT signaling in T cells through CNR2. Signal Transduct Target Ther. 2022;7(1):99. Published 2022 Apr 6. doi:10.1038/s41392-022-00918-y(IF:18.187)

[2] Zhang Z, Chen C, Yang F, et al. Itaconate is a lysosomal inducer that promotes antibacterial innate immunity [published online ahead of print, 2022 May 25]. Mol Cell. 2022;S1097-2765(22)00443-9. doi:10.1016/j.molcel.2022.05.009(IF:17.970)

[3] Cai J, Peng J, Zang X, et al. Mammary Leukocyte-Assisted Nanoparticle Transport Enhances Targeted Milk Trace Mineral Delivery [published online ahead of print, 2022 Jun 30]. Adv Sci (Weinh). 2022;e2200841. doi:10.1002/advs.202200841(IF:17.521)

[4] Jin H, Liu K, Tang J, et al. Genetic fate-mapping reveals surface accumulation but not deep organ invasion of pleural and peritoneal cavity macrophages following injury. Nat Commun. 2021;12(1):2863. Published 2021 May 17. doi:10.1038/s41467-021-23197-7(IF:14.919)

[5] Sheng D, Ma W, Zhang R, et al. Ccl3 enhances docetaxel chemosensitivity in breast cancer by triggering proinflammatory macrophage polarization [published correction appears in J Immunother Cancer. 2022 Jun;10(6):]. J Immunother Cancer. 2022;10(5):e003793. doi:10.1136/jitc-2021-003793(IF:13.751)

[6] Zhao L, Zhang H, Liu X, et al. TGR5 deficiency activates antitumor immunity in non-small cell lung cancer via restraining M2 macrophage polarization. Acta Pharm Sin B. 2022;12(2):787-800. doi:10.1016/j.apsb.2021.07.011(IF:11.614)

[7] Xia L, Zhang C, Lv N, et al. AdMSC-derived exosomes alleviate acute lung injury via transferring mitochondrial component to improve homeostasis of alveolar macrophages. Theranostics. 2022;12(6):2928-2947. Published 2022 Mar 21. doi:10.7150/thno.69533(IF:11.556)

[8] Zhang X, Hou L, Li F, et al. Piezo1-mediated mechanosensation in bone marrow macrophages promotes vascular niche regeneration after irradiation injury. Theranostics. 2022;12(4):1621-1638. Published 2022 Jan 16. doi:10.7150/thno.64963(IF:11.556)

[9] Wang H, Li L, Li Y, et al. Intravital imaging of interactions between iNKT and kupffer cells to clear free lipids during steatohepatitis. Theranostics. 2021;11(5):2149-2169. Published 2021 Jan 1. doi:10.7150/thno.51369(IF:11.556)

[10] Xun J, Du L, Gao R, et al. Cancer-derived exosomal miR-138-5p modulates polarization of tumor-associated macrophages through inhibition of KDM6B. Theranostics. 2021;11(14):6847-6859. Published 2021 May 3. doi:10.7150/thno.51864(IF:11.556)

[11] Zuo L, Li J, Zhang X, et al. Aberrant mesenteric adipose extracellular matrix remodeling is involved in adipocyte dysfunction in Crohn's disease: The role of TLR-4-mediated macrophages [published online ahead of print, 2022 Jun 16]. J Crohns Colitis. 2022;jjac087. doi:10.1093/ecco-jcc/jjac087(IF:9.071)

[12] Huang C, Wang J, Liu H, et al. Ketone body β-hydroxybutyrate ameliorates colitis by promoting M2 macrophage polarization through the STAT6-dependent signaling pathway. BMC Med. 2022;20(1):148. Published 2022 Apr 15. doi:10.1186/s12916-022-02352-x(IF:8.775)

[13] Jiang P, Gao W, Ma T, et al. CD137 promotes bone metastasis of breast cancer by enhancing the migration and osteoclast differentiation of monocytes/macrophages. Theranostics. 2019;9(10):2950-2966. Published 2019 May 9. doi:10.7150/thno.29617(IF:8.063)

[14] Yang XL, Wang G, Xie JY, et al. The Intestinal Microbiome Primes Host Innate Immunity against Enteric Virus Systemic Infection through Type I Interferon. mBio. 2021;12(3):e00366-21. Published 2021 May 11. doi:10.1128/mBio.00366-21(IF:7.867)

[15] Sun Z, Huang W, Zheng Y, et al. Fpr2/CXCL1/2 Controls Rapid Neutrophil Infiltration to Inhibit Streptococcus agalactiae Infection. Front Immunol. 2021;12:786602. Published 2021 Nov 24. doi:10.3389/fimmu.2021.786602(IF:7.561)

[16] Cai J, Cui X, Wang X, You L, Ji C, Cao Y. A Novel Anti-Infective Peptide BCCY-1 With Immunomodulatory Activities. Front Immunol. 2021;12:713960. Published 2021 Jul 22. doi:10.3389/fimmu.2021.713960(IF:7.561)