DOI: 10.5061/dryad.s4mw6m9cj
title: Data from: Comparison of osteoclast differentiation protocols from human induced pluripotent stem cells of different tissue origins

description: Background: Ever since their discovery, induced pluripotent stem cells
 (iPSCs) have been extensively differentiated into a large variety of cell
 types. However, a limited amount of work has been dedicated to
 differentiating iPSCs into osteoclasts. While several differentiation
 protocols have been published, it remains unclear which protocols or
 differentiation methods are preferable regarding the differentiation of
 osteoclasts. Methods: In this study, we compare the osteoclastogenesis
 capacity of a peripheral blood mononuclear cell (PBMC)-derived iPSC line
 to a fibroblast-derived iPSC line in conjunction with either embryoid
 body-based or monolayer-based differentiation strategies. Both cell lines
 and differentiation protocols were investigated regarding their ability to
 generate osteoclasts and their inherent robustness and ease of use. The
 ability of both cell lines to remain undifferentiated while propagating
 using a feeder-free system was assessed using alkaline phosphatase
 staining. This was followed by evaluating mesodermal differentiation and
 the characterization of hematopoietic progenitor cells using flow
 cytometry. Finally, osteoclast yield and functionality based on resorptive
 activity, Cathepsin K, and tartrate-resistant acid phosphatase (TRAP)
 expression were assessed. Results were validated using qRT-PCR throughout
 the differentiation stages. Results: Embryoid-body-based differentiation
 yielded CD45+, CD14+, and CD11b+ subpopulations, which in turn
 differentiated into osteoclasts which demonstrated TRAP positivity,
 Cathepsin K expression, and mineral resorptive capabilities. This was
 regardless of which iPSC line was used. Monolayer-based differentiation
 yielded lower quantities of hematopoietic cells that were mostly CD34+ and
 did not subsequently differentiate into osteoclasts. Conclusions: The
 outcome of this study demonstrates the successful differentiation of
 osteoclasts from iPSCs in conjunction with the embryoid-based
 differentiation method, while the monolayer-based method did not yield
 osteoclasts. No differences were observed regarding osteoclast
 differentiation between the PBMC and fibroblast-derived iPSC lines.
description: # Data from: Comparison of osteoclast differentiation protocols from human
 induced pluripotent stem cells of different tissue origins
 [https://doi.org/10.5061/dryad.s4mw6m9cj](https://doi.org/10.5061/dryad.s4mw6m9cj) The data made available in this data set support the conclusions made in the publication in Stem Cell Research & Therapy entitled "Comparison of osteoclast differentiation protocols from human induced pluripotent stem cells of different tissue origins". ## Description of the data and file structure The data provided in this dataset support the conclusions of each figure in the publication. Hence, the dataset is structured according to the figure numbers of the publication. Zip files are entitled according to the figure number of the publication (Figure 2 Data, Figure 3 Data, etc.). Files are named to be self-explanatory. Zip files related to figures that include graphs in the publications (Figure 5, Figure 7 and Figure 8) contain Excel files showing the raw data and GraphPad Prism files demonstrating the graphs used in the figure. The Zip file corresponding to Figure 4 contains a FlowJo workbook with the flow cytometry data, as well as the raw .fcs files. Flow data is structured into 2 groups (Fibroblast-iPSC and PBMC-iPSC) with 3 subgroups each (EB, MB and undifferentiated iPSCs as neg. control). Each subgroup has 14 tubes. Tube 1 is negative control with no fluorochromes, tubes 2-7 are isotype controls, tubes 8-13 are fluorescence minus one controls (FMO) and cells tube 14 are stained with the full antibody panel. Fibroblast-derived iPSC are sometimes referred to as "GM28404” or “Fibroblast-iPSC”, PBMC-derived iPSC are sometimes referred to "MCND-S2" or “PBMC-iPSC” within the dataset. Additionally, the embryoid-body based differentiatoin method (EB) is sometimes referred to as "Rössler" while the monolayer-based method (MB) is sometimes referred to as "STEMdiff". ## Code/Software As mentioned above, flow cytometry data is stored in a FlowJo workbook file, raw quantitative data is provided in Excel sheets, graphs used in figures are stored in GraphPad Prism files.