TY - JOUR

T1 - Single-cell high content imaging parameters predict functional phenotype of cultured human bone marrow stromal stem cells

AU - Kowal, Justyna M.

AU - Schmal, Hagen

AU - Halekoh, Ulrich

AU - Hjelmborg, Jacob B.

AU - Kassem, Moustapha

PY - 2020

Y1 - 2020

N2 - Cultured human bone marrow stromal (mesenchymal) stem cells (hBM-MSCs) are heterogenous cell populations exhibiting variable biological properties. Quantitative high-content imaging technology allows identification of morphological markers at a single cell resolution that are determinant for cellular functions. We determined the morphological characteristics of cultured primary hBM-MSCs and examined their predictive value for hBM-MSC functionality. BM-MSCs were isolated from 56 donors and characterized for their proliferative and differentiation potential. We correlated these data with cellular and nuclear morphological features determined by Operetta; a high-content imaging system. Cell area, cell- and nucleus geometry of cultured hBM-MSCs exhibited significant correlation with expression of hBM-MSC membrane markers: ALP, CD146, CD271. Proliferation capacity correlated negatively with cell and nucleus area and positively with cytoskeleton texture features. In addition, in vitro differentiation to osteoblasts as well as in vivo heterotopic bone formation was associated with decreased ratio of nucleus width to length. Multivariable analysis applying a stability selection procedure identified nuclear geometry and texture as predictors for hBM-MSCs differentiation potential to osteoblasts or adipocytes. Our data demonstrate that by employing a limited number of cell morphological characteristics, it is possible to predict the functional phenotype of cultured hBM-MSCs and thus can be used as a screening test for “quality” of hBM-MSCs prior their use in clinical protocols. Stem Cells Translational Medicine 2019.

AB - Cultured human bone marrow stromal (mesenchymal) stem cells (hBM-MSCs) are heterogenous cell populations exhibiting variable biological properties. Quantitative high-content imaging technology allows identification of morphological markers at a single cell resolution that are determinant for cellular functions. We determined the morphological characteristics of cultured primary hBM-MSCs and examined their predictive value for hBM-MSC functionality. BM-MSCs were isolated from 56 donors and characterized for their proliferative and differentiation potential. We correlated these data with cellular and nuclear morphological features determined by Operetta; a high-content imaging system. Cell area, cell- and nucleus geometry of cultured hBM-MSCs exhibited significant correlation with expression of hBM-MSC membrane markers: ALP, CD146, CD271. Proliferation capacity correlated negatively with cell and nucleus area and positively with cytoskeleton texture features. In addition, in vitro differentiation to osteoblasts as well as in vivo heterotopic bone formation was associated with decreased ratio of nucleus width to length. Multivariable analysis applying a stability selection procedure identified nuclear geometry and texture as predictors for hBM-MSCs differentiation potential to osteoblasts or adipocytes. Our data demonstrate that by employing a limited number of cell morphological characteristics, it is possible to predict the functional phenotype of cultured hBM-MSCs and thus can be used as a screening test for “quality” of hBM-MSCs prior their use in clinical protocols. Stem Cells Translational Medicine 2019.

KW - cell and nucleus morphology

KW - high-content imaging

KW - human stromal/mesenchymal stem cells

KW - osteoblastic and adipocytic differentiation

KW - proliferation

U2 - 10.1002/sctm.19-0171

DO - 10.1002/sctm.19-0171

M3 - Journal article

C2 - 31758755

AN - SCOPUS:85075436742

VL - 9

SP - 189

EP - 202

JO - Stem cells translational medicine

JF - Stem cells translational medicine

SN - 2157-6564

IS - 2

ER -