Transplantation of whole kidney marrow in adult zebrafish

Jocelyn LeBlanc, Teresa V. Bowman, Leonard Zon

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Hematopoietic stem cells (HSC) are a rare population of pluripotent cells that maintain all the differentiated blood lineages throughout the life of an organism. The functional definition of a HSC is a transplanted cell that has the ability to reconstitute all the blood lineages of an irradiated recipient long term. This designation was established by decades of seminal work in mammalian systems. Using hematopoietic cell transplantation (HCT) and reverse genetic manipulations in the mouse, the underlying regulatory factors of HSC biology are beginning to be unveiled, but are still largely under-explored. Recently, the zebrafish has emerged as a powerful genetic model to study vertebrate hematopoiesis. Establishing HCT in zebrafish will allow scientists to utilize the large-scale genetic and chemical screening methodologies available in zebrafish to reveal novel mechanisms underlying HSC regulation. In this article, we demonstrate a method to perform HCT in adult zebrafish. We show the dissection and preparation of zebrafish whole kidney marrow, the site of adult hematopoiesis in the zebrafish, and the introduction of these donor cells into the circulation of irradiated recipient fish via intracardiac injection. Additionally, we describe the post-transplant care of fish in an "ICU" to increase their long-term health. In general, gentle care of the fish before, during, and after the transplant is critical to increase the number of fish that will survive more than one month following the procedure, which is essential for assessment of long term (<3 month) engraftment. The experimental data used to establish this protocol will be published elsewhere. The establishment of this protocol will allow for the merger of large-scale zebrafish genetics and transplant biology.

Original languageEnglish (US)
Article numbere159
JournalJournal of Visualized Experiments
Issue number2
DOIs
StatePublished - Dec 2006
Externally publishedYes

Fingerprint

Zebrafish
Stem cells
Kidney Transplantation
Fish
Transplants
Bone Marrow
Hematopoietic Stem Cells
Cell Transplantation
Fishes
Blood
Cytology
Dissection
Intensive care units
Hematopoiesis
Screening
Reverse Genetics
Cells
Health
Genetic Models
Genetic Testing

Keywords

  • Developmental biology
  • HSC
  • Issue 2
  • Stem cells
  • Transplant
  • Zebrafish

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Transplantation of whole kidney marrow in adult zebrafish. / LeBlanc, Jocelyn; Bowman, Teresa V.; Zon, Leonard.

In: Journal of Visualized Experiments, No. 2, e159, 12.2006.

Research output: Contribution to journalArticle

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