99mTc-methyl-diphosphonate binding to mineral deposits in cultures of marrow-derived mesenchymal stem cells in osteogenic medium

Tobias Grossner, Tobias Gotterbarm, Victor H. Gerbaudo, Uwe Haberkorn, Myron Spector

Research output: Contribution to journalArticle

Abstract

Osteogenic differentiation of adult mesenchymal stem cells (MSCs) is a promising method for the therapy of critical size bone defects and other applications, drawing attention to the importance of in vitro assays for its evaluation. While there are standardized protocols to induce osteogenic differentiation in monolayer cultures, there is a lack of validated nondestructive procedures to quantify the osteogenic potential of MSCs directly by accessing the mineral deposition. In this study, we present a new method to determine the osteogenic potential of MSCs using the radioactive tracer, 99mTechnetium-methylene diphosphonate (99mTc-MDP), to bind in vitro to newly produced mineral deposits from osteogenic-induced stem cells. Thirty five-millimeter Petri dishes were seeded with goat and human MSCs and differentiated into the osteogenic lineage. 99mTc-MDP was applied to each dish, and acquisition of the bound tracer was imaged with a gamma camera. The results revealed a significantly higher uptake in the osteogenic-differentiated MSCs for both goat and human cells (p < 0.008 goat; 0.01 human), compared with controls grown in expansion medium. We were able to show that 99mTc-MDP labeling is a suitable method to quantify the amount of extracellular mineral deposited by MSCs. Goat and human MSCs displayed similar uptake characteristics (Spearman-Rho Correlation coefficient r = 0.603). The radionuclide method was validated by quantitative Alizarin Red staining, which showed a high correlation (Spearman-Rho Correlation coefficient r = 0.668). This radionuclide method opens a new approach for evaluating osteogenic potential of MSCs and for longitudinal studies of the process. The work is notable for describing a highly sensitive, quantitative, and nondestructive method for evaluating the in vitro amount of mineral accompanying different types of osteogenic differentiation of mesenchymal stem cells in a monolayer cell culture. What is so unique and useful about the method is that it has the potential to be used to define the kinetics of the differentiation process, reflected in the mineralization, without destroying the monolayer. Therefore, it remains intact for further experiments.

Original languageEnglish (US)
Pages (from-to)49-57
Number of pages9
JournalTissue Engineering - Part C: Methods
Volume25
Issue number1
DOIs
StatePublished - Jan 1 2019

Fingerprint

Mineral resources
Diphosphonates
Stem cells
Mesenchymal Stromal Cells
Cell culture
Minerals
Bone Marrow
Goats
Monolayers
Radioisotopes
Radioactive Tracers
Alizarin
Adult Stem Cells
Gamma Cameras
Radioactive tracers
Longitudinal Studies
Labeling
Stem Cells
Cell Culture Techniques
Assays

Keywords

  • mesenchymal stem cells
  • mineral quantification
  • osteogenic differentiation
  • technetium

ASJC Scopus subject areas

  • Bioengineering
  • Medicine (miscellaneous)
  • Biomedical Engineering

Cite this

99mTc-methyl-diphosphonate binding to mineral deposits in cultures of marrow-derived mesenchymal stem cells in osteogenic medium. / Grossner, Tobias; Gotterbarm, Tobias; Gerbaudo, Victor H.; Haberkorn, Uwe; Spector, Myron.

In: Tissue Engineering - Part C: Methods, Vol. 25, No. 1, 01.01.2019, p. 49-57.

Research output: Contribution to journalArticle

Grossner, Tobias ; Gotterbarm, Tobias ; Gerbaudo, Victor H. ; Haberkorn, Uwe ; Spector, Myron. / 99mTc-methyl-diphosphonate binding to mineral deposits in cultures of marrow-derived mesenchymal stem cells in osteogenic medium. In: Tissue Engineering - Part C: Methods. 2019 ; Vol. 25, No. 1. pp. 49-57.
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