Specificity of 3D MSC Spheroids Microenvironment: Impact on MSC Behavior and Properties

Jauković, Aleksandra; Abadjieva, Desislava; Trivanović, Drenka; Stoyanova, Elena; Kostadinova, Milena; Pashova, Shina; Kestendjieva, Snejana; Кукољ, Тамара; Ješeta, Michal; Kistanova, Elena; Mourdjeva, Milena

doi:10.1007/s12015-020-10006-9

Cited by 68 publications

(47 citation statements)

References 152 publications

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“…These observations are consistent with changes in cell survival. In the majority of published experiments, 3D culture is a transient stage which last usually up to three div, no longer than seven div-especially when it comes to the use of low-attachment surface [4] among others because of poor MSC survival in 3D conditions. In our experiments 3D culture also reduced cell viability.…”

Section: Discussionmentioning

confidence: 99%

“…Monolayer culture system allows MSC to attach to the surface just like in natural environment and to expand in two dimensions. In spite of its widespread, this method has multiple limitations, and it is discussed how close is to the natural cell environment of MSC [3,4].…”

Section: Introductionmentioning

confidence: 99%

“…Except analysis of neural phenotype, other aspects such as proliferation, senescence and stemness were not so broadly taken into consideration during research. Moreover, majority of MSC spheroids experiments were a short-term-3D cultured did not last up to seven days in vitro of culture (div) [4] and results were obtained usually during first three div. There were little evidences whether observed effects were transient or constant whether how cells would react for prolonged 3D conditions.…”

Section: Introductionmentioning

confidence: 99%

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Effect of Long-Term 3D Spheroid Culture on WJ-MSC

Kamińska

Wędzińska

Kot

et al. 2021

Cells

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The aim of our work was to develop a protocol enabling a derivation of mesenchymal stem/stromal cell (MSC) subpopulation with increased expression of pluripotent and neural genes. For this purpose we used a 3D spheroid culture system optimal for neural stem cells propagation. Although 2D culture conditions are typical and characteristic for MSC, under special treatment these cells can be cultured for a short time in 3D conditions. We examined the effects of prolonged 3D spheroid culture on MSC in hope to select cells with primitive features. Wharton Jelly derived MSC (WJ-MSC) were cultured in 3D neurosphere induction medium for about 20 days in vitro. Then, cells were transported to 2D conditions and confront to the initial population and population constantly cultured in 2D. 3D spheroids culture of WJ-MSC resulted in increased senescence, decreased stemness and proliferation. However long-termed 3D spheroid culture allowed for selection of cells exhibiting increased expression of early neural and SSEA4 markers what might indicate the survival of cell subpopulation with unique features.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of Long-Term 3D Spheroid Culture on WJ-MSC

Kamińska

Wędzińska

Kot

et al. 2021

Cells

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“…To address the limitations associated with conventional adherent cultures and facilitate scaffold-free cell transplantation, a new culture system of MSCs using ultra-low-attachment culture dishes was recently developed to obtain MSC spheroids, which were referred to as "mesenspheres" (Mendez-Ferrer et al, 2010;Baraniak and McDevitt, 2012;Isern et al, 2013). MSC spheroids are considered as a promising tool for regenerative medicine (Jaukovic et al, 2020). The culture dishes in these studies had a predetermined microwell size at the bottom, which resulted in mesenspheres with a fixed size.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, we determined the number of spheroids and spheroid size, instead of generating a growth curve. MSC spheroids have a unique microenvironment compared with adherent MSCs, such as metabolic change or epigenetic regulation (Jaukovic et al, 2020). Other reports have shown that engineering MSC spheroids potentiated cell-cell and cell-ECM interaction (Lee et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

A Shaking-Culture Method for Generating Bone Marrow Derived Mesenchymal Stromal/Stem Cell-Spheroids With Enhanced Multipotency in vitro

Niibe

Ohori-Morita

Zhang

et al. 2020

Front. Bioeng. Biotechnol.

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Mesenchymal stromal/stem cells (MSCs), which generally expand into adherent monolayers, readily lose their proliferative and multilineage potential following repeated passages. Floating culture systems can be used to generate MSC spheroids, which are expected to overcome limitations associated with conventional adherent cultures while facilitating scaffold-free cell transplantation. However, the phenotypic characteristics of spheroids after long-term culture are unknown. In addition, regenerative therapies require new culture systems to maintain their undifferentiated state. In this study, we established a novel culture method employing three-dimensional (3D) "shaking" to generate MSC spheroids using bone marrow derived MSCs. Floating 3D cultures of mouse or human MSCs formed spheroids after shaking (85-95 rpm), within 1 month. These spheroids maintained their osteogenic-, adipogenic-, and chondrogenicdifferentiation capacity. The adipogenic-differentiation capacity of adherent cultured mouse and human MSCs, which is lost following several passages, was remarkedly restored by shaking-culture. Notably, human MSC spheroids exhibited a renewable "undifferentiated MSC-pool" property, wherein undifferentiated MSCs grew from spheroids seeded repeatedly on a plastic culture dish. These data suggest that the shaking-culture method maintains and restores multipotency that is lost following monolayer expansion and thereby shows potential as a promising strategy for regenerative therapies with mesenchymal tissues.

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3D Stem Cell Spheroids with 2D Hetero‐Nanostructures for In Vivo Osteogenic and Immunologic Modulated Bone Repair

Liu,

Astudillo Potes,

Dashtdar

et al. 2024

Adv Healthcare Materials

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Three‐dimensional (3D) stem cell spheroids have immense potential for various tissue engineering applications. However, current spheroid fabrication techniques encounter cell viability issues due to limited oxygen access for cells trapped within the core, as well as non‐specific differentiation issues due to the complicated environment following transplantation. In this study, we developed functional 3D spheroids using mesenchymal stem cells (MSCs) with two‐dimensional (2D) hetero‐nanostructures (HNS) composed of single‐stranded DNA (ssDNA) binding carbon nanotubes (sdCNT) and gelatin‐bind black phosphorus nanosheets (gBPNS). An osteogenic molecule, dexamethasone (DEX), was further loaded to fabricate an sdCNTgBP‐DEX hetero‐nanostructure. This approach aimed to establish a multi‐functional cell‐inductive 3D spheroid with improved oxygen transportation through hollow nanotubes, stimulated stem cell growth by phosphate ions supplied from BP oxidation, in‐situ immunoregulation, and osteogenesis induction by DEX molecules after implantation. Initial transplantation of the 3D spheroids in rat calvarial bone defect showed in vivo macrophage shifts to an M2 phenotype, leading to a pro‐healing microenvironment for regeneration. Prolonged implantation demonstrated outstanding in vivo neovascularization, osteointegration, and new bone regeneration. Therefore, these engineered 3D spheroids hold great promise for bone repair as they allow for stem cell delivery and provide immunoregulative and osteogenic signals within an all‐in‐one construct.This article is protected by copyright. All rights reserved

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Specificity of 3D MSC Spheroids Microenvironment: Impact on MSC Behavior and Properties

Cited by 68 publications

References 152 publications

Effect of Long-Term 3D Spheroid Culture on WJ-MSC

Effect of Long-Term 3D Spheroid Culture on WJ-MSC

A Shaking-Culture Method for Generating Bone Marrow Derived Mesenchymal Stromal/Stem Cell-Spheroids With Enhanced Multipotency in vitro

3D Stem Cell Spheroids with 2D Hetero‐Nanostructures for In Vivo Osteogenic and Immunologic Modulated Bone Repair

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