Cell death, cavitation and spontaneous multi‐differentiation of dental pulp stem cells‐derived spheroids <i>in vitro</i>: A journey to survival and organogenesis

Xiao, Li; Kumazawa, Yasuo; Okamura, Hisashi

doi:10.1111/boc.201400024

Cited by 25 publications

(17 citation statements)

References 48 publications

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“…Enhanced expression of Oct 4 and Nanog in sphe roids of human dental pulp stem cells compared to monolayer culture has been revealed with PCR (Xiao et al, 2014). Cells in spheroids obtained from human placenta MSCs underwent epigenetic changes that facilitated increased expression of the pluripotent genes Oct 4, Nanog, and Sox 2 (Guo et al, 2014).…”

Section: Surface Marker Expression In Fetmsc and Fetmsc And Their Sphmentioning

confidence: 99%

“…It has been demonstrated that MSC spheroids exhibit increased anti inflammation effect, increased differentiation potential, and enhanced expression of cytokines and pluripotent genes. It facilitates the understanding of organogenesis mechanisms and their application in the regenerative medicine (Alimperti et al, 2014;Bogdanova Jatniece et al, 2014;Guo et al, 2014;Murphy et al, 2014;Xiao et al, 2014;Li et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Cellular spheroids obtained from mesenchymal stem cells derived from bone marrow and limb muscle of early human embryo

et al. 2015

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Cellular spheroids were obtained from mesenchymal stem cells derived from different tissues of 5 to 6 week old embryos: bone marrow (FetMSC) and limb muscle (M FetMSC). The properties of these cells maintained in monolayer (2D cultivation) or spheroids (3D cultivation) were compared. Cellular sphe roids were obtained from monolayer cultures at the sixth passage after decryopreservation and assayed 48 h after their formation. Unlike monolayer cultures, spheroids are heterogeneous cell populations consisted of fibroblast like and epithelioid cells. Two day old spheroids are actively proliferating structures. Cell surface markers were analyzed by flow cytometry. Monolayer culture and cellular spheroids expressed CD44, CD73, CD90, CD105, and HLA ABC common for mesenchymal stem cells (MSCs) and lacked CD34 and HLA DR. However, expression of CD90 and CD105 antigens was significantly lower in spheroids than in mono layer cultures. Expression of transcription factors and surface antigens typical for human embryonic stem cells were analyzed with immunofluorescence and flow cytometry. Expression of Sox 2 and SSEA 4 in 2D and 3D cultures, lack of Oct 4 in 2D cultures, and its increase in 3D cultures were found. Both monolayer cells and spheroids were able to differentiate into osteogenic, chondrogenic, and adipogenic lineages, although monolayer cultures and spheroids differed. Adipogenic differentiation was more active in the cellular spheroids from M FetMSC cells compared to corresponding monolayer cultures. Differences between the 2D and 3D cultures of both lines have been shown by the character of chondrogenic differentiation. As a whole, these findings confirm the MSC status for the cellular spheroids derived from FetMSC and M FetMSC mono layer cultures and attest to their increased differentiation potential compared to monolayer cultures.Abbreviations: ECM-extracellular matrix, MSC-mesenchy mal stem cell, ESC-embryonic stem cell, 2D-two dimen sional culture system, 3D-three dimensional culture system. 432 CELL AND TISSUE BIOLOGY Vol. 9No. 6 2015 KRYLOVA et al.

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Section: Surface Marker Expression In Fetmsc and Fetmsc And Their Sphmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cellular spheroids obtained from mesenchymal stem cells derived from bone marrow and limb muscle of early human embryo

et al. 2015

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“…Spheroid models have been developed for in vitro studies in developmental science and stem cell research (Xiao & Tsutsui 2013, Xiao et al 2014. Also in vitro biocompatibility and toxicity assays have been established based on these culture models (Banerjee & Bhonde 2006.…”

Section: Introductionmentioning

confidence: 99%

Formation of spheroids by dental pulp cells in the presence of hypoxia and hypoxia mimetic agents

et al. 2017

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Aim To evaluate the impact of hypoxia and hypoxia mimetic agents (HMA) on the formation and activity of spheroids by dental pulp cells (DPC). Methodology DPC on agarose-coated plates were treated with hypoxia and the HMA dimethyloxallyl glycine (DMOG), desferrioxamine (DFO) and L-mimosine (L-MIM). Images of spheroids were taken directly after seeding and at 6 h and 24 h. Spheroid sizes were quantified by area measurement with ImageJ software. Viability was assessed with Live-Dead staining, MTT and resazurin-based toxicity assay. Production of VEGF, IL-8 and SDF-1 was evaluated using immunoassays. Data were analysed using KruskalWallis test and post hoc Mann-Whitney U-test. Results DPC formed spheroids in the presence of hypoxia, HMA and combined treatment with hypoxia and HMA. No pronounced difference in spheroid size was found in the groups treated with hypoxia, DMOG, DFO, L-MIM and the combination of hypoxia and the HMA relative to their normoxic controls (P > 0.05). Spheroids appeared vital in Live-Dead and MTT staining and the resazurin-based toxicity assay. Evaluation of protein production with immunoassays revealed significantly enhanced levels of VEGF and IL-8 (P < 0.05), but there was no significant effect on SDF-1 production (P > 0.05). Treatment with a combination of hypoxia and HMA did not further boost VEGF and IL-8 production (P > 0.05). Conclusions Pre-conditioning with hypoxia and HMA increased the pro-angiogenic capacity of spheroids whilst not interfering with their formation. Preclinical studies will reveal whether pre-conditioning of spheroids with hypoxia and HMA can effectively improve the efficiency of cell transplantation approaches for regenerative endodontics.

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“…found that dermal papilla (DP) spheroids of larger size displayed slightly decreased viability on D5 55 . A large spheroid core is often deficient in oxygen and/or nutrients and shows excessive accumulation of catabolites and low pH 56, 57 . We found that CECs in spheroid culture displayed enhanced expression of the corneal endothelial marker ATP1A1.…”

Section: Discussionmentioning

confidence: 99%

In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers

Han

Lei

et al. 2017

Sci Rep

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Corneal endothelial cells (CECs) are very important for the maintenance of corneal transparency. However, in vitro, CECs display limited proliferation and loss of phenotype via endothelial to mesenchymal transformation (EMT) and cellular senescence. In this study, we demonstrate that continuous supplementary nutrition using a perfusion culture bioreactor and three-dimensional (3D) spheroid culture can be used to improve CEC expansion in culture and to construct a tissue-engineered CEC layer. Compared with static culture, perfusion-derived CECs exhibited an increased proliferative ability as well as formed close cell-cell contact junctions and numerous surface microvilli. We also demonstrated that the CEC spheroid culture significantly down-regulated gene expression of the proliferation marker Ki67 and EMT-related markers Vimentin and α-SMA, whereas the gene expression level of the CEC marker ATP1A1 was significantly up-regulated. Furthermore, use of the perfusion system in conjunction with a spheroid culture on decellularized corneal scaffolds and collagen sheets promoted the generation of CEC monolayers as well as neo-synthesized ECM formation. This study also confirmed that a CEC spheroid culture on a curved collagen sheet with controlled physiological intraocular pressure could generate a CEC monolayer. Thus, our results show that the use of a perfusion system and 3D spheroid culture can promote CEC expansion and the construction of tissue-engineered corneal endothelial layers in vitro.

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Cell death, cavitation and spontaneous multi‐differentiation of dental pulp stem cells‐derived spheroids in vitro: A journey to survival and organogenesis

Abstract: DPSC-derived spheroids can mimic the developmental process of cell survival, cavitation and spontaneous multi-differentiation on matrigel under certain conditions. This work allows for functional studies to investigate organ regeneration with human DPSCs in vitro.

Cited by 25 publications

References 48 publications

Cellular spheroids obtained from mesenchymal stem cells derived from bone marrow and limb muscle of early human embryo

Cellular spheroids obtained from mesenchymal stem cells derived from bone marrow and limb muscle of early human embryo

Formation of spheroids by dental pulp cells in the presence of hypoxia and hypoxia mimetic agents

In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers

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