Intravital longitudinal wide-area imaging of dynamic bone marrow engraftment and multilineage differentiation through nuclear-cytoplasmic labeling

PLoS One. 2017 Nov 3;12(11):e0187660. doi: 10.1371/journal.pone.0187660. eCollection 2017.

Abstract

Bone marrow is a vital tissue that produces the majority of erythrocytes, thrombocytes, and immune cells. Bone marrow transplantation (BMT) has been widely performed in patients with blood disorders and cancers. However, the cellular-level behaviors of the transplanted bone marrow cells over wide-areas of the host bone marrow after the BMT are not fully understood yet. In this work, we performed a longitudinal wide-area cellular-level observation of the calvarial bone marrow after the BMT in vivo. Using a H2B-GFP/β-actin-DsRed double-transgenic mouse model as a donor, a subcellular-level nuclear-cytoplasmic visualization of the transplanted bone marrow cells was achieved, which enabled a direct in vivo dynamic monitoring of the distribution and proliferation of the transplanted bone marrow cells. The same spots in the wide-area of the calvarial bone marrow were repeatedly identified using fluorescently labeled vasculature as a distinct landmark. It revealed various dynamic cellular-level behaviors of the transplanted BM cells in early stage such as cluster formation, migration, and active proliferation in vivo.

MeSH terms

  • Animals
  • Bone Marrow / diagnostic imaging
  • Bone Marrow / pathology*
  • Bone Marrow Transplantation*
  • Cell Differentiation*
  • Cell Lineage
  • Cell Nucleus / metabolism*
  • Cytoplasm / metabolism*
  • Flow Cytometry
  • Humans
  • Longitudinal Studies
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Confocal

Grants and funding

This work was supported by the Global Frontier Project (NRF-2013M3A6A4044716) and the Bio & Medical Technology Development Program (NRF-2017M3A9E4047243) funded by the Ministry of Science, ICT and Future Planning, Republic of Korea and the Korea Healthcare Technology R&D Project (HI13C2181, HI15C0399) funded by Ministry of Health and Welfare, Republic of Korea.