Analysis of Spontaneous and Induced Osteogenic Differentiation in 3D-micromasses of Human Multipotent Stem Cells

Julian Lommen; MICHAEL SUS; Karin Berr; Norbert R. Kübler; Fabian Langenbach; Christoph Sproll; Max Wilkat; Felix Schrader; Jörg Handschel; Lara Schorn

doi:10.21873/invivo.12804

January 1, 2022In VivoOpen Access

Analysis of Spontaneous and Induced Osteogenic Differentiation in 3D-micromasses of Human Multipotent Stem Cells

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Key Points

To assess how a dexamethasone, ascorbic acid, and β-glycerophosphate (DAG) cocktail, along with MAPK, ERK, and PI3K/Akt pathway inhibitors, affects the growth and mineralization of human unrestricted somatic stem cell (USSC) micromasses.
Cultured three human USSC cell lines (USSC-18, USSC-8, and USSC-8/17) as 3D micromasses in standard medium.
Induced osteogenic differentiation using DAG (50 μM dexamethasone, 50 mM ascorbic acid, and 1 M β-glycerolphosphate) with or without signaling pathway inhibitors FR180204, LY294002, or U0126.
Stained microtome slices using OsteoImage and evaluated micromass size and mineralization using fluorescence microscopy.
DAG treatment produced a significant increase in micromass size and mineralization on day 3 (p<0.001), day 6 (p<0.001), and day 7 (p<0.001).
The ERK inhibitor FR180204 reduced growth and mineralization in non-DAG-treated cells (p<0.001) but enhanced mineralization in DAG-treated cells (p=0.014), whereas the MAPK inhibitor U0126 reduced both growth and mineralization in DAG-treated cells (p=0.001).
The PI3K/Akt inhibitor LY294002 did not significantly alter micromass growth but significantly suppressed mineralization (p<0.001).

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Implication

In vitro study demonstrates that osteogenic supplements and signaling inhibitors regulate mineralization in stem cell micromasses, suggesting targets to improve bone tissue engineering.

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Cite This Study

Lommen et al. (2022) studied this question.

synapsesocial.com/papers/6a9ddac78d7c9026f618abc4 https://doi.org/10.21873/invivo.12804

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Analysis of Spontaneous and Induced Osteogenic Differentiation in 3D-micromasses of Human Multipotent Stem Cells

Key Points

No abstract yet

Implication

Discussion

Cite This Study

Analysis of Spontaneous and Induced Osteogenic Differentiation in 3D-micromasses of Human Multipotent Stem Cells

Key Points

No abstract yet

Implication

Discussion

Cite This Study

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