Characterization of a pluripotent stem cell-derived matrix with powerful osteoregenerative capabilities

McNeill, Eoin P.; Zeitouni, Suzanne; Pan, Simin; Haskell, Andrew W.; Cesarek, Michael; Tahan, Daniel; Clough, Bret H.; Krause, U.; Dobson, Lauren K.; García, Mayra; Kung, Christopher; Zhao, Qingguo; Saunders, W. B.; Liu, Fei; Kaunas, Roland; Gregory, Carl A.

doi:10.1038/s41467-020-16646-2

Cited by 43 publications

(52 citation statements)

References 70 publications

(117 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The iPSC-MSCs were recovered from frozen vials of a cell bank that has been characterized extensively including the trilineage differentiation in comparison with bone marrow MSCs from multiple donors [ 16 , 19 ]. These cells were plated at a density of 500 cells per square centimeter of growth area in 17% FBS αMEM medium at 37 °C and 5% CO 2 and passaged upon 70–80% confluence.…”

Section: Methodsmentioning

confidence: 99%

“…These iPSC-derived MSCs (iPSC-MSCs) are highly consistent in the homing capacities to various cancers and the expression of surface molecules related to cancer targeting [ 16 – 18 ]. The osteogenic potentials of our iPSC-MSCs are superior to BM-MSCs [ 16 , 19 ], while the immunomodulatory capacities are comparable between these two types of MSCs [ 20 , 21 ]. We have prepared nanoghosts from membrane-only ghost iPSC-MSCs and nanovesicles from intact iPSC-MSCs respectively by serial extrusions [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Extracellular vesicle mimics made from iPS cell-derived mesenchymal stem cells improve the treatment of metastatic prostate cancer

et al. 2021

Self Cite

View full text Add to dashboard Cite

Background Extracellular vesicles (EVs) and their mimics from mesenchymal stem cells (MSCs) are promising drug carriers to improve cancer treatment, but their application is hindered by donor variations and expansion limitations of conventional tissue-derived MSCs. To circumvent these issues, we made EV-mimicking nanovesicles from standardized MSCs derived from human induced pluripotent stem cells (iPSCs) with a theoretically limitless expandability, and examined the targeting capacity of these nanovesicles to prostate cancer. Methods Nanovesicles are made from intact iPSC-MSCs through serial extrusion. The selective uptake of fluorescently labeled nanovesicles by prostate cancer cells vs. non-tumor cells was examined with flow cytometry. For in vivo tracing, nanovesicles were labeled with fluorescent dye DiR or renilla luciferase. In mice carrying subcutaneous or bone metastatic PC3 prostate cancer, the biodistribution of systemically infused nanovesicles was examined with in vivo and ex vivo imaging of DiR and luminescent signals. A chemotherapeutic drug, docetaxel, was loaded into nanovesicles during extrusion. The cytotoxicities of nanovesicle-encapsulated docetaxel on docetaxel-sensitive and -resistant prostate cancer cells and non-tumor cells were examined in comparison with free docetaxel. Therapeutic effects of nanovesicle-encapsulated docetaxel were examined in mice carrying subcutaneous or bone metastatic prostate cancer by monitoring tumor growth in comparison with free docetaxel. Results iPSC-MSC nanovesicles are more selectively taken up by prostate cancer cells vs. non-tumor cells in vitro compared with EVs, membrane-only EV-mimetic nanoghosts and liposomes, which is not affected by storage for up to 6 weeks. In mouse models of subcutaneous and bone metastatic PC3 prostate cancer, systemically infused nanovesicles accumulate in tumor regions with significantly higher selectivity than liposomes. The loading of docetaxel into nanovesicles was efficient and did not affect the selective uptake of nanovesicles by prostate cancer cells. The cytotoxicities of nanovesicle-encapsulated docetaxel are significantly stronger on docetaxel-resistant prostate cancer cells and weaker on non-tumor cells than free docetaxel. In mouse models of subcutaneous and bone metastatic prostate cancer, nanovesicle-encapsulated docetaxel significantly decreased the tumor growth and toxicity to white blood cells compared with free docetaxel. Conclusions Our data indicate that EV-mimicking iPSC-MSC nanovesicles are promising to improve the treatment of metastatic prostate cancer.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Extracellular vesicle mimics made from iPS cell-derived mesenchymal stem cells improve the treatment of metastatic prostate cancer

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…Each sample was dissolved upon shaking, and the supernatant was collected after 15 min. The collected supernatant was measured for absorbance at 405 nm using a spectrometer (McNeill et al, 2020), and the amount of calcification was quantitatively compared with sampling n 3.…”

Section: Alizarin Red S Staining Methodsmentioning

confidence: 99%

Osteogenic Response to Polysaccharide Nanogel Sheets of Human Fibroblasts After Conversion Into Functional Osteoblasts by Direct Phenotypic Cell Reprogramming

Nakai

Yamamoto

Kishida

et al. 2021

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Human dermal fibroblasts (HDFs) were converted into osteoblasts using a ALK inhibitor II (inhibitor of transforming growth factor-β signal) on freeze-dried nanogel-cross-linked porous (FD-NanoClip) polysaccharide sheets or fibers. Then, the ability of these directly converted osteoblasts (dOBs) to produce calcified substrates and the expression of osteoblast genes were analyzed in comparison with osteoblasts converted by exactly the same procedure but seeded onto a conventional atelocollagen scaffold. dOBs exposed to FD-NanoClip in both sheet and fiber morphologies produced a significantly higher concentration of calcium deposits as compared to a control cell sample (i.e., unconverted fibroblasts), while there was no statistically significant difference in calcification level between dOBs exposed to atelocollagen sheets and the control group. The observed differences in osteogenic behaviors were interpreted according to Raman spectroscopic analyses comparing different polysaccharide scaffolds and Fourier transform infrared spectroscopy analyses of dOB cultures. This study substantiates a possible new path to repair large bone defects through a simplified transplantation procedure using FD-NanoClip sheets with better osteogenic outputs as compared to the existing atelocollagen scaffolding material.

show abstract

“…Therefore, a clear need exists for the development of an effective technology with efficacy similar to autografting but without the associated limitations. The most feasible form for this technology is a cell-scaffold composite; however, achieving clinical efficacy, reproducibility, biocompatibility, and an effective production pipeline have hampered development of this strategy ( McNeill et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Canine Mesenchymal Stromal Cell-Mediated Bone Regeneration is Enhanced in the Presence of Sub-Therapeutic Concentrations of BMP-2 in a Murine Calvarial Defect Model

Dobson

Zeitouni

McNeill

et al. 2021

Front. Bioeng. Biotechnol.

Self Cite

View full text Add to dashboard Cite

Novel bone regeneration strategies often show promise in rodent models yet are unable to successfully translate to clinical therapy. Sheep, goats, and dogs are used as translational models in preparation for human clinical trials. While human MSCs (hMSCs) undergo osteogenesis in response to well-defined protocols, canine MSCs (cMSCs) are more incompletely characterized. Prior work suggests that cMSCs require additional agonists such as IGF-1, NELL-1, or BMP-2 to undergo robust osteogenic differentiation in vitro. When compared directly to hMSCs, cMSCs perform poorly in vivo. Thus, from both mechanistic and clinical perspectives, cMSC and hMSC-mediated bone regeneration may differ. The objectives of this study were twofold. The first was to determine if previous in vitro findings regarding cMSC osteogenesis were substantiated in vivo using an established murine calvarial defect model. The second was to assess in vitro ALP activity and endogenous BMP-2 gene expression in both canine and human MSCs. Calvarial defects (4 mm) were treated with cMSCs, sub-therapeutic BMP-2, or the combination of cMSCs and sub-therapeutic BMP-2. At 28 days, while there was increased healing in defects treated with cMSCs, defects treated with cMSCs and BMP-2 exhibited the greatest degree of bone healing as determined by quantitative μCT and histology. Using species-specific qPCR, cMSCs were not detected in relevant numbers 10 days after implantation, suggesting that bone healing was mediated by anabolic cMSC or ECM-driven cues and not via engraftment of cMSCs. In support of this finding, defects treated with cMSC + BMP-2 exhibited robust deposition of Collagens I, III, and VI using immunofluorescence. Importantly, cMSCs exhibited minimal ALP activity unless cultured in the presence of BMP-2 and did not express endogenous canine BMP-2 under any condition. In contrast, human MSCs exhibited robust ALP activity in all conditions and expressed human BMP-2 when cultured in control and osteoinduction media. This is the first in vivo study in support of previous in vitro findings regarding cMSC osteogenesis, namely that cMSCs require additional agonists to initiate robust osteogenesis. These findings are highly relevant to translational cell-based bone healing studies and represent an important finding for the field of canine MSC-mediated bone regeneration.

show abstract

Characterization of a pluripotent stem cell-derived matrix with powerful osteoregenerative capabilities

Cited by 43 publications

References 70 publications

Extracellular vesicle mimics made from iPS cell-derived mesenchymal stem cells improve the treatment of metastatic prostate cancer

Extracellular vesicle mimics made from iPS cell-derived mesenchymal stem cells improve the treatment of metastatic prostate cancer

Osteogenic Response to Polysaccharide Nanogel Sheets of Human Fibroblasts After Conversion Into Functional Osteoblasts by Direct Phenotypic Cell Reprogramming

Canine Mesenchymal Stromal Cell-Mediated Bone Regeneration is Enhanced in the Presence of Sub-Therapeutic Concentrations of BMP-2 in a Murine Calvarial Defect Model

Contact Info

Product

Resources

About