Capacity of the medullary cavity of tibia and femur for intra-bone marrow transplantation in mice


Autoři: Dieter Fink aff001;  Ulrike Pfeiffenberger aff001;  Tina Bernthaler aff001;  Sophie Schober aff001;  Kerstin E. Thonhauser aff001;  Thomas Rülicke aff001
Působiště autorů: Institute of Laboratory Animal Science, Department of Biomedical Sciences, University of Veterinary Medicine Vienna, Vienna, Austria aff001;  Ludwig Boltzmann Institute for Hematology and Oncology, Medical University of Vienna, Austria aff002
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224576

Souhrn

Intra-bone marrow transplantation (IBMT) has been adapted for mouse models to improve the seeding efficiency of transplanted hematopoietic stem and progenitor cells. Commonly used injection volumes for IBMT into the tibia differ between 10 and 40 μL even though considerable amounts of injected cells leak into the blood circulation immediately after injection. Injection of 3 μL trypan blue into the tibia of dead BALB/c mice showed staining in large vessels of hind limbs, even without supporting circulation. We therefore tested the effective capacity of the medullary cavity of dissected tibiae and femora of different mouse strains by bioluminescence imaging after injection of luciferase expressing cells. Cell leakage was already observed at 3 μL of injection volume and the measured emission rate increased significantly when 5 and 10 μL of volume with the same cell concentration were injected. Surprisingly, increasing injection volumes containing constant cell amounts resulted in comparable emission rates, suggesting a similar amount of leaked and absorbed cells independent of the injection volume. However, the absorption of a specific amount of injected cells could not be confirmed, as the ratio of leaked to absorbed cells was similar between IBMT that were performed with a constant injection volume containing either low or high cell amounts. In summary, for optimal cell transplantation via IBMT in mice we suggest to inject a high concentrated cell suspension with a maximum injection volume of 3 μL.

Klíčová slova:

Bioluminescence – Bone marrow cells – Bone marrow transplantation – Luciferase – Mouse models – Hematopoietic stem cell transplantation


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Článek vyšel v časopise

PLOS One


2019 Číslo 11