Quantification of bone marrow interstitial pH and calcium concentration by intravital ratiometric imaging

S-C. A. Yeh, J. Hou, J. W. Wu, S. Yu, Y. Zhang, K. D. Belfield, F. D. Camargo and C. P. Lin ()
Additional contact information
S-C. A. Yeh: Harvard Medical School
J. Hou: Harvard Medical School
J. W. Wu: Harvard Medical School
S. Yu: New Jersey Institute of Technology
Y. Zhang: New Jersey Institute of Technology
K. D. Belfield: New Jersey Institute of Technology
F. D. Camargo: Boston Children’s Hospital
C. P. Lin: Harvard Medical School

Nature Communications, 2022, vol. 13, issue 1, 1-13

Abstract: Abstract The fate of hematopoietic stem cells (HSCs) can be directed by microenvironmental factors including extracellular calcium ion concentration ([Ca2+]e), but the local [Ca2+]e around individual HSCs in vivo remains unknown. Here we develop intravital ratiometric analyses to quantify the absolute pH and [Ca2+]e in the mouse calvarial bone marrow, taking into account the pH sensitivity of the calcium probe and the wavelength-dependent optical loss through bone. Unexpectedly, the mean [Ca2+]e in the bone marrow (1.0 ± 0.54 mM) is not significantly different from the blood serum, but the HSCs are found in locations with elevated local [Ca2+]e (1.5 ± 0.57 mM). With aging, a significant increase in [Ca2+]e is found in M-type cavities that exclusively support clonal expansion of activated HSCs. This work thus establishes a tool to investigate [Ca2+]e and pH in the HSC niche with high spatial resolution and can be broadly applied to other tissue types.

Date: 2022
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DOI: 10.1038/s41467-022-27973-x

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