Comparison of haematopoietic stem cell engraftment through the retro-orbital venous sinus and the lateral vein: alternative routes for bone marrow transplantation in mice

Abstract

Bone marrow transplantation in mice is performed by intravenous administration of haematopoietic repopulating cells, usually via the lateral tail vein. This technique can be technically challenging to carry out and may cause distress to the mice. The retro-orbital sinus is a large area where there is a confluence of several vessels that provides an alternative route for intravenous access. Retro-orbital injection, although aesthetically unpleasant, can be performed rapidly without requiring mechanical restriction or heat-induced vasodilation. In addition, this technique can be easily learned by novice manipulators. This route of administration has been reported for use in bone marrow transplantation but there is no comparison of retro-orbital and tail vein injections reported for this specific purpose, although both routes have been compared for many other applications. Here, we provide for the first time a comprehensive comparison between tail vein and retro-orbital injections for two different bone marrow transplant scenarios in P3B and B6D2F1 mice. In both cases, no significant differences regarding donor engraftment were observed between mice transplanted using each of the techniques. Haematological counts and leukocyte subpopulation distribution were practically identical between both animal groups. Moreover, donor engraftment levels were less homogenous when cells were transplanted by tail vein injection, probably due to a higher risk of failure associated with this technique. All these data suggest that retro-orbital injection is a compelling alternative to conventional tail vein injection for bone marrow transplant in mice, providing similar and more homogenous haematopoietic reconstitution.

Off-Target and Tumor-Specific Accumulation of Monocytes, Macrophages and Myeloid-Derived Suppressor Cells after Systemic Injection

“…Labelled cells were gently vortexed prior to injection with a 29G insulin syringe. Unless noted otherwise, 100 μl of cells suspended in DPBS were injected in the right orbital plexus of anesthetized mice at a concentration of 10x10 6 /ml [21] , [22] , [23] , [24] . Local injection occurred through intratumoral injection of 10 5 DiR-labelled monocytes (in 50 μl DPBS) into 10-days old 4T1 tumors.…”

Study logistics that can impact medical countermeasure efficacy testing in mouse models of radiation injury

“…Additionally, retro-orbital injections are less stressful on the mice and are easier to perform in darker pigmented mice where tail veins are harder to visualize. Studies comparing hematopoietic stem cell engraftment ( Leon-Rico et al 2015 ), tumor uptake ( Kim et al 2010 ) and therapeutic agents ( Steel et al 2008 ) administered through the retro-orbital venous sinus and lateral tail vein showed no significant difference between the two methods, providing compelling evidence for the use of retro-orbital injections for administration of cellular therapies. It is important to note that irradiated mice, with a compromised hematopoietic system and fragile vasculature, may be more susceptible than unirradiated mice to differences between these methods, thus rigorous training of study personnel and pilot studies are warranted to determine which route is most efficacious in radiation experiments.…”

Assessment of human platelet survival in the NOD/SCID mouse model: technical considerations

“…However, owing to the assumption that PLTs would become activated from shear stress in the narrow tail vein, the model had been established using the retro‐orbital plexus for PLT infusion, resulting in 8% to 15% human PLTs out of all circulating PLTs 30 minutes after injection. Although retro‐orbital injection has been shown to be an efficient route to transfuse non‐cellular fluids or stem cells, the injection of highly concentrated PLTs in human plasma seems to be different. In this study we did not perform a systematic evaluation of the approach‐associated mortality.…”