Bone Marrow Transplantation Procedures in Mice to Study Clonal Hematopoiesis

Eunbee Park; Megan  A. Evans; Heather Doviak; Keita Horitani; Hayato Ogawa; Yoshimitsu Yura; Ying Wang; Soichi Sano; Kenneth Walsh

doi:10.3791/61875

Bone Marrow Transplantation Procedures in Mice to Study Clonal Hematopoiesis

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Bone Marrow Transplantation Procedures in Mice to Study Clonal Hematopoiesis

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08:00 min

May 26, 2021

DOI: 10.3791/61875-v

Eunbee Park¹, Megan A. Evans⁴, Heather Doviak⁴, Keita Horitani⁴, Hayato Ogawa⁴, Yoshimitsu Yura⁴, Ying Wang², Soichi Sano^3,4, Kenneth Walsh^1,4

¹Department of Biochemistry and Molecular Genetics,University of Virginia School of Medicine, ²Department of Cardiology, Xinqiao Hospital,Army Medical University, ³Department of Cardiology,Osaka City University Graduate School of Medicine, ⁴Hematovascular Biology Center, Robert M. Berne Cardiovascular Research Center,University of Virginia School of Medicine

Overview

This article describes three methods of bone marrow transplantation (BMT) used to study clonal hematopoiesis in mouse models. The methods include BMT with total-body irradiation, shielded irradiation, and adoptive BMT without pre-conditioning.

Key Study Components

Area of Science

Bone marrow transplantation
Clonal hematopoiesis
Mouse models

Background

Bone marrow transplantation is critical for studying hematopoiesis.
Total body irradiation can negatively impact cardiovascular health.
Alternative methods aim to reduce side effects associated with traditional BMT.
Understanding these methods is essential for improving experimental outcomes.

Purpose of Study

To evaluate the physiological effects of different BMT methods.
To assess how these methods influence clonal hematopoiesis.
To provide protocols for minimizing adverse effects during BMT.

Methods Used

BMT with total-body irradiation
BMT with shielded irradiation
Adoptive BMT without pre-conditioning
Detailed procedural demonstrations by lab members

Main Results

Demonstrated the impact of total body radiation on cardiovascular organs.
Outlined procedures for alternative BMT methods.
Provided insights into the effects on disease pathogenesis.
Showcased the importance of shielding during irradiation.

Conclusions

Alternative BMT methods can mitigate side effects of total body irradiation.
Understanding these methods is crucial for future research in clonal hematopoiesis.
Protocols established can guide researchers in experimental setups.

Frequently Asked Questions

What are the three BMT methods discussed?

The methods are BMT with total-body irradiation, shielded irradiation, and adoptive BMT without pre-conditioning.

Why is total-body irradiation used?

It is traditionally used to prepare the recipient for bone marrow transplantation.

What are the potential side effects of total-body irradiation?

It can negatively impact cardiovascular organs and alter disease pathogenesis.

How do alternative BMT methods help?

They minimize or avoid the side effects associated with total-body irradiation.

Who demonstrates the BMT procedures?

Eunbee Park, a graduate student, and Megan Evans, a postdoctoral fellow, demonstrate the procedures.

What is the significance of shielding during irradiation?

Shielding helps protect vital organs from radiation exposure during the procedure.

We describe three methods of bone marrow transplantation (BMT): BMT with total-body irradiation, BMT with shielded irradiation, and BMT method with no pre-conditioning (adoptive BMT) for the study of clonal hematopoiesis in mouse models.

Our protocol will help researchers understand the physiological effects of three different bone marrow transplantation methods and how they affect experimental outcomes in clonal hematopoiesis setting. Total body radiation bone marrow transplantation can negatively impact cardiovascular organs and alter disease pathogenesis. Thus, our lab has developed two alternative methods to minimize or avoid possible side-effects.

Demonstrating the procedures will be Eunbee Park a graduate student, and Megan Evans a postdoctoral fellow, both from my laboratory For thorax and abdomen shielding, place the adjustable tray and the x-ray irradiator at the correct distance to achieve uniform irradiation. And place the anesthetized mice on a flat led plate inverted to each other in the supine position. Secure the paws to the plate with tape, and place the led shielding so that the lower end aligns with the xiphisternum bone and the upper end of the lead shield sits near the thymus.

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