Intravital microscopy and statistical image analysis of early engraftment of haematopoietic stem cells reveal dynamic and evolving cellular behaviours linked to progressive differentiation.
- 1Imperial College London
Abstract
Haematopoietic stem cells (HSCs) have long been used in the clinic for bone marrow transplantation applications, critical for the survival of an increasingly wide range of patients with haematological, oncological and immunological pathologies. Despite this, little is known about the mechanisms through which relatively few stem cells are able to regenerate the entire haematopoietic tissue of transplant recipients. To gain insights on this biological process, we used intravital microscopy of calvarium bone marrow, collecting tissue-wide images and a total of 850 hours of tracks of engrafting haematopoietic stem and progenitor cells from 24 hours to 8 days following injection in lethally irradiated recipients. Analysis of the data revealed that regenerating HSCs and their immediate progeny are highly dynamic, migrating through the parenchyma at both the microscopic and near-macroscopic scales, i.e. within and in-and-out of fields of view. HSC-derived cell clusters, expanded locally between day 2 and day 4 post-transplant, leading to patches of densely populated bone marrow by day 8. Single cell level analysis highlighted increasingly heterogenous cellular behaviours over time. Track clustering based on migration and niche interaction parameters combined with post-tracking whole mount immunostaining revealed that persistence in the vicinity of nestin-GFP perivascular cells and relatively slow movement correlated with stemness, while more heterogeneous niche interactions coupled with slowest or faster migration correlated with differentiation, validated through flow cytometry analysis and functional studies. The findings presented here shed light on the fundamental principles driving haematopoietic regeneration in transplantation settings.
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BibTeX
@misc{anton2026intravital,
title = {{Intravital microscopy and statistical image analysis of early engraftment of haematopoietic stem cells reveal dynamic and evolving cellular behaviours linked to progressive differentiation.}},
author = {S. Gonzalez Anton and C. Liu and R. A. Khorshed and M. C. Romero-Mulero and G. Adams and B. Partridge and Z. A. Lu and N. Cabezas-Wallscheid and C. Pospori and C. F. Lee and K. R. Duffy and C. Lo Celso},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.11.750961},
url = {https://www.biorxiv.org/content/10.64898/2026.09.11.750961v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Intravital microscopy and statistical image analysis of early engraftment of haematopoietic stem cells reveal dynamic and evolving cellular behaviours linked to progressive differentiation.
AU - S. Gonzalez Anton
AU - C. Liu
AU - R. A. Khorshed
AU - M. C. Romero-Mulero
AU - G. Adams
AU - B. Partridge
AU - Z. A. Lu
AU - N. Cabezas-Wallscheid
AU - C. Pospori
AU - C. F. Lee
AU - K. R. Duffy
AU - C. Lo Celso
PY - 2026
DA - 2026/09/11
PB - bioRxiv
DO - 10.64898/2026.09.11.750961
UR - https://www.biorxiv.org/content/10.64898/2026.09.11.750961v1
AB - Haematopoietic stem cells (HSCs) have long been used in the clinic for bone marrow transplantation applications, critical for the survival of an increasingly wide range of patients with haematological, oncological and immunological pathologies. Despite this, little is known about the mechanisms through which relatively few stem cells are able to regenerate the entire haematopoietic tissue of transplant recipients. To gain insights on this biological process, we used intravital microscopy of calvarium bone marrow, collecting tissue-wide images and a total of 850 hours of tracks of engrafting haematopoietic stem and progenitor cells from 24 hours to 8 days following injection in lethally irradiated recipients. Analysis of the data revealed that regenerating HSCs and their immediate progeny are highly dynamic, migrating through the parenchyma at both the microscopic and near-macroscopic scales, i.e. within and in-and-out of fields of view. HSC-derived cell clusters, expanded locally between day 2 and day 4 post-transplant, leading to patches of densely populated bone marrow by day 8. Single cell level analysis highlighted increasingly heterogenous cellular behaviours over time. Track clustering based on migration and niche interaction parameters combined with post-tracking whole mount immunostaining revealed that persistence in the vicinity of nestin-GFP perivascular cells and relatively slow movement correlated with stemness, while more heterogeneous niche interactions coupled with slowest or faster migration correlated with differentiation, validated through flow cytometry analysis and functional studies. The findings presented here shed light on the fundamental principles driving haematopoietic regeneration in transplantation settings.
ER -