Induction of mesenchymal stem cell differentiation by co-culturing with mature cells in double-layered 2-methacryloyloxyethyl phosphorylcholine polymer hydrogel matrices

Ishihara, Kazuhíko; Kaneyasu, Miu; Fukazawa, Kyoko; Zhang, Ren; Teramura, Yuji

doi:10.1039/d1tb01817e

Cited by 4 publications

(3 citation statements)

References 42 publications

(74 reference statements)

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“…This study compared the structural features of Ti membranes between the single-layered 2-dimensional (2D) culture and multi-layered 3D-like scaffold and observed the proliferation of the C3H10T1/2 cells, a mouse MSC-like progenitor cell line [ 18 ], under both conditions.…”

Section: Introductionmentioning

confidence: 99%

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

et al. 2023

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In vitro studies on adherent cells require a process of passage to dissociate the cells from the culture substrate using enzymes or other chemical agents to maintain cellular activity. However, these proteolytic enzymes have a negative influence on the viability and phenotype of cells. The mesenchymal stem cell (MSC)-like cell line, C3H10T1/2, adhered, migrated, and proliferated to the same extent on newly designed microporous titanium (Ti) membrane and conventional culture dish, and spontaneous transfer to another substrate without enzymatic or chemical dissociation was achieved. The present study pierced a 10 μm-thick pure Ti sheet with 25 μm square holes at 75 μm intervals to create a dense porous structure with biomimetic topography. The pathway of machined holes allowed the cells to access both sides of the membrane frequently. In a culture with Ti membranes stacked above- and below-seeded cells, cell migration between the neighboring membranes was confirmed using the through-holes of the membrane and contact between the membranes as migration routes. Furthermore, the cells on each membrane migrated onto the conventional culture vessel. Therefore, a cell culture system with enzyme-free passaging was developed.

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Section: Introductionmentioning

confidence: 99%

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

et al. 2023

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“…5 On the other hand, isolated adult stem cells may inadvertently differentiate into undesirable cell lineages during in vitro continuously culturing in conventional tissue culture plates (TCP), 5 leading to the loss of their normal phenotype. 10 Indeed, biochemical and biophysical cues in the surrounding environment influence cell proliferation and differentiation, 11 and regulate the MSCs' differentiation ability into a specific lineage. 4 The in vivo microenvironment provides numerous regulating stimuli not available in conventional cell culture methods.…”

Section: Introductionmentioning

confidence: 99%

Molecular imprinting as a simple way for the long-term maintenance of the stemness and proliferation potential of adipose-derived stem cells: an in vitro study

et al. 2022

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“…MPC-based polymers have shown excellent bio-and hemocompatibility and have been used as coating materials for various medical implants such as contact lenses, guidewires, ear grommets, and coronary stents. [26][27][28][29] MPC 30 -DEA 70 (MPC refers to 2-(methacryloyloxy)-ethyl phosphorylcholine and DEA refers to 2-(diethylamino)-ethyl methacrylates; 30 and 70 refer to the mean degrees of polymerization of each block) is a weak cationic diblock copolymer, whose highlight is the main structural feature of solution complexation between the copolymers and antisense oligodeoxynucleotides (AS-ODNs). [30][31][32] MPC segments are zwitterionic and strongly hydrated, and they provide hydrophilic affinity for the polyplexes to remain soluble.…”

Section: Introductionmentioning

confidence: 99%

The complexing of cationic copolymer MPC₃₀-DEA₇₀with TGF-β1 antisense oligodeoxynucleotide and transfection into cardiomyocytesin vitro

Agyekum

Zhang

et al. 2022

J Biomater Appl

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Although gene therapy is an attractive option for the treatment of cardiovascular diseases, the ideal gene delivery systems are still under investigation and must meet the following criteria: safety, adequate gene transfer efficiency, and stable expression of the transgene for a duration appropriate for treating the disease. In this study, we developed a cationic phosphorylcholine-containing diblock copolymer, namely MPC30-DEA70, as carrier systems to deliver a chemically synthesized transforming growth factor-beta 1(TGF-β1) antisense oligonucleotide (AS-ODN) into cardiomyocytes (CMs) to observe the cell transfection efficiency of MPC30-DEA70 and the inhibition effect on the expression of TGF-β1. MPC30-DEA70/TGF-β1 AS-ODN complexes were formed through complexation between copolymer MPC30-DEA70 (N) and AS-ODN (P) at different N/P ratios and were characterized by DNA electrophoresis. Notably, the cytotoxicity and cell growth inhibition assay showed that the MPC30-DEA70 had low cytotoxicity to CMs within the effective transfection dosage range (<20 μL/mL). CLSM/TEM images displayed that most of the AS-ODN molecules engulfed by cells were located around the cell nuclei, and a few entered into the cell nuclei without harming the organelles in the cell. Transfection studies from CMs indicated a steady increase of transfection efficiency with increasing N/P ratios. The expression levels of TGF-β1 mRNA and protein in CMs were significantly inhibited at high N/P ratios. This study shows that MPC30-DEA70 can function as an effective transgenic vector into CMs and that TGF-β1 AS-ODN delivered by MPC30-DEA70 can silence the expression of the TGF-β1 gene efficiently and specifically and thereafter antagonize TGF-β1-mediated biological function in cardiomyocytes.

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Induction of mesenchymal stem cell differentiation by co-culturing with mature cells in double-layered 2-methacryloyloxyethyl phosphorylcholine polymer hydrogel matrices

Abstract: The effects of differentiated cells on stem cell differentiation were analyzed by co-culture using a cell-encapsulated double-layered hydrogel system. As a polymer hydrogel matrix, a water-soluble zwitterionic polymer having both...

Cited by 4 publications

References 42 publications

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

Molecular imprinting as a simple way for the long-term maintenance of the stemness and proliferation potential of adipose-derived stem cells: an in vitro study

The complexing of cationic copolymer MPC₃₀-DEA₇₀with TGF-β1 antisense oligodeoxynucleotide and transfection into cardiomyocytesin vitro

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Induction of mesenchymal stem cell differentiation by co-culturing with mature cells in double-layered 2-methacryloyloxyethyl phosphorylcholine polymer hydrogel matrices

Abstract: The effects of differentiated cells on stem cell differentiation were analyzed by co-culture using a cell-encapsulated double-layered hydrogel system. As a polymer hydrogel matrix, a water-soluble zwitterionic polymer having both...

Cited by 4 publications

References 42 publications

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

Molecular imprinting as a simple way for the long-term maintenance of the stemness and proliferation potential of adipose-derived stem cells: an in vitro study

The complexing of cationic copolymer MPC30-DEA70with TGF-β1 antisense oligodeoxynucleotide and transfection into cardiomyocytesin vitro

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The complexing of cationic copolymer MPC₃₀-DEA₇₀with TGF-β1 antisense oligodeoxynucleotide and transfection into cardiomyocytesin vitro