In vivo flow cytometry combined with intravital microscopy to monitor kinetics of transplanted bone marrow mononuclear cells in peripheral blood and bone marrow

Mol Biol Rep. 2020 Jan;47(1):1-10. doi: 10.1007/s11033-019-04608-x. Epub 2019 Dec 7.

Abstract

Bone marrow mononuclear cells (BM-MNCs) transplantation has evolved as a promising experimental treatment in various regenerative therapy fields, especially in clinical hematopoietic stem cells transplantation (HSCT). In vitro methods have mainly been used to study the pre-clinical kinetics of BM-MNCs in mice after transplantation. And it is difficult to monitor the dynamic homing of BM-MNCs in living mice. The present study obtained the kinetics of transplanted BM-MNCs in the peripheral blood (PB) and the dynamic homing of BM-MNCs in the BM in living mice by a combination of in vivo flow cytometry (IVFC) and calvarium intravital microscopy. We found out that BM-MNCs were cleared rapidly from the PB and mainly localized to various hematopoietic tissues after transplantation. The number of BM-MNCs in the PB decreased over time accompanied by an increase in the BM indeed after transplantation. In addition, a lower number of BM-MNCs were found home to calvaria than long bone, probably indicating long bone marrow might also be an important hematopoietic organ. Clinical studies will benefit from non-invasive measurements to monitor the dynamic homing of transplanted cells. Our pre-clinical kinetics of BM-MNCs in living mice will have important clinical guiding significance in HSCT and other regenerative therapy fields.

Keywords: Bone marrow mononuclear cells; Hematopoietic stem cell transplantation; Homing; In vivo; Stem cell.

MeSH terms

  • Animals
  • Blood Cells / cytology
  • Bone Marrow / metabolism
  • Bone Marrow Cells / physiology*
  • Bone Marrow Transplantation*
  • Cell Movement*
  • Cell Tracking / methods*
  • Chemotaxis, Leukocyte / physiology
  • Flow Cytometry / methods*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Hindlimb
  • Intravital Microscopy / methods*
  • Kinetics
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Neovascularization, Physiologic

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins