In vivo pharmacology

Drug dosing

We provide dosing services by Intravenous (IV), Intraperitoneal (IP), Per Os (PO), subcutaneous (SQ), Intradermal (ID), under the foot pad injection or by other customized administration. We also provide Alzet osmotic pump implantation services. Osmotic pumps are miniature infusion pumps which provide a safe and effective method for continuous and controlled agent delivery in vivo. The pumps function through an osmotic gradient between the pump and the tissue where the pump is implanted. Osmotic pumps can be implanted subcutaneously, and can also be used for targeted delivery in blood, CSF, brain, or other tissues. Pumps and solutions to be infused must be prepared sterile and must be implanted using aseptic sterile technique, with the animal under general anesthesia.


MTD and ADMET studies

Maximum Tolerated Dose (MTD) studies, pharmacokinetic (PK), pharmacodynamics (PD), and ADMET(absorption, distribution, metabolism, excretion, and toxicity) studies are essential for the development of novel therapeutic compounds or combination therapies. We offer various dosing services followed by timed blood and organ collection to generate MTD/PK/PD data at the convenience of the researchers.


Blood collection 

Serum preparation

Collect whole blood in regular tubes, leave on ice to coagulate, and centrifuge at 1600g at 40C for 15 min, to result in serum. EDTA tubes should be avoided if NMR spectroscopy is going to be used. 

[Olaf Beckonert, H. C. (2007). Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts. Nature Protocol, 2: 2692-2703.]

 

Plasma preparation

Collect whole blood in heparin coated tubes (BD Microtainer® tube with Lithium Heparin additive BD Microgard™, green cap) or EDTA tubes (BD Microtainer® tube with Dipotassium EDTA, Beadless additive BD Microgard™, lavender cap) and spin down at 8,000rpm at 40C for 10 min in a desktop centrifuge. Collect supernatant in regular Eppendorf tubes and store in -80oC immediately. 

[S D Christensen, L. F. (2009). Quality of plasma sampled by different methods for multiple blood sampling in mice. Laboratory Animals, 43: 65-71.]

 

Retro-orbital collection: Mice are anesthetized with isoflurane. A capillary tube will be inserted at the lateral canthus, and the sinus is punctured with gentle pressure and twisting motion. Blood is collected at the tip of the glass capillary tube.

Facial vein collection:  A lancet is used to puncture the vessel at a small area on the ventral jaw. Blood is collected directly into the appropriate tube.

Distal tail collection: Mice will be warmed by a heating lamp for 5 min. A scalpel will be used to slice gently at tail vein. Blood is collected directly into the appropriate tube.

[Jonathan Hoggatt, A. F. (2016). Bleeding the laboratory mouse: Not all methods are equal. Experimental Hematology, 44: 132-137.]


Efficacy study

After tumors are implanted, compounds are treated and their efficacy is monitored by measuring tumor growth. 

The figure on the right shows results from a tumor xenograft experiment testing the efficacy of H2 agonist cimetidine in inhibiting the growth of A549 lung adenoma cell line tumors in SCID mice. Three treatment groups (25, 50, or 100 mg/kg per injection) and one control group (PBS) were used. Another group was treated with doxorubicin as a positive control.

[Marina Sirota, et al. (2011). Discovery and Preclinical Validation of Drug Indications Using Compendia of Public Gene Expression Data. Science Translational Medicine, 3: 96ra77.]


Irradiation

We provide whole body irradiation for murine models.

Here we show results from one of our irradiation and nanoparticle combination studies. Arrows show time line (days) of administration of nanoparticle (NP) and radiation (Gy) treatment.  

 

 

The results show in vivo nanoparticle therapy with single intratumoural injection. Human non-small cell lung cancer xenografts were established by injecting A459 cells subcutaneously into the right hindlimb of a SCID mouse. Tumors were grown for one week prior to treatment. Mice were shielded from radiation except for the exposed tumour bearing flank. Mice were either irradiated with 0 Gy or 2 Gy, daily for five days. NP, nanoparticle, IR, irradiation. 

 

[Helen E. Townley, et al. (2012). In vivo demonstration of enhanced radiotherapy using rare earth doped titania nanoparticles. Nanoscale, 5043–5050.]


Summary of in vivo pharmacology services

1.    Drug dosing

2.    MTD and ADMET

3.    Blood collection and preparation

4.    Efficacy study

5.    Irradiation