Protocol for noninvasive imaging of hematopoietic reconstitution in live mice using Akaluc bioluminescence

Area of Science:

• Hematology
• Biomedical Imaging
• Stem Cell Biology

Background:

• Hematopoietic stem cell (HSC) transplantation is crucial for treating blood disorders.
• Monitoring hematopoietic reconstitution noninvasively is essential for assessing transplant success.
• Current methods often lack spatiotemporal resolution or are invasive.

Purpose of the Study:

• To develop and present a protocol for noninvasive, spatiotemporal visualization of hematopoietic reconstitution in live mice.
• To enable quantitative in vivo tracking of HSC engraftment and recovery dynamics.

Main Methods:

• Akaluc gene transduction into murine HSCs.
• Ex vivo expansion of hematopoietic stem and progenitor cells.
• Longitudinal bioluminescence imaging post transplantation in live mice.

Main Results:

• Demonstrated successful spatiotemporal visualization of hematopoietic recovery in spleen and bone marrow.
• Illustrated quantitative in vivo tracking of hematopoietic reconstitution dynamics.
• Validated the protocol for whole-body resolution following lethal irradiation.

Conclusions:

• The developed Akaluc-based bioluminescence imaging (AkaBLI) protocol provides a powerful tool for studying hematopoietic reconstitution.
• This method allows for noninvasive, quantitative, and longitudinal assessment of stem cell engraftment and recovery.
• The protocol facilitates a deeper understanding of hematopoietic reconstitution dynamics in vivo.