Novel method to produce a layered 3D scaffold for human pluripotent stem cell-derived neuronal cells

Honkamäki, Laura; Joki, Tiina; Grigoryev, Nikita A.; Levón, Kalle; Ylä‐Outinen, Laura; Narkilahti, Susanna

doi:10.1016/j.jneumeth.2020.109043

Cited by 11 publications

(10 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The immunostaining protocol was adapted from Honkamäki et al (2021) and performed on 2–4 technical replicates for each cell donor. After 6 days of culture microvascular networks were washed with DPBS and fixed with 4% paraformaldehyde (Sigma-Aldrich) in DPBS for 30 min at RT following by three washes with DPBS.…”

Section: Methodsmentioning

confidence: 99%

Vasculogenic Potency of Bone Marrow- and Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells Results in Differing Vascular Network Phenotypes in a Microfluidic Chip

Mykuliak

Yrjänäinen

Mäki

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

The vasculature is an essential, physiological element in virtually all human tissues. Formation of perfusable vasculature is therefore crucial for reliable tissue modeling. Three-dimensional vascular networks can be formed through the co-culture of endothelial cells (ECs) with stromal cells embedded in hydrogel. Mesenchymal stem/stromal cells (MSCs) derived from bone marrow (BMSCs) and adipose tissue (ASCs) are an attractive choice as stromal cells due to their natural perivascular localization and ability to support formation of mature and stable microvessels in vitro. So far, BMSCs and ASCs have been compared as vasculature-supporting cells in static cultures. In this study, BMSCs and ASCs were co-cultured with endothelial cells in a fibrin hydrogel in a perfusable microfluidic chip. We demonstrated that using MSCs of different origin resulted in vascular networks with distinct phenotypes. Both types of MSCs supported formation of mature and interconnected microvascular networks-on-a-chip. However, BMSCs induced formation of fully perfusable microvasculature with larger vessel area and length whereas ASCs resulted in partially perfusable microvascular networks. Immunostainings revealed that BMSCs outperformed ASCs in pericytic characteristics. Moreover, co-culture with BMSCs resulted in significantly higher expression levels of endothelial and pericyte-specific genes, as well as genes involved in vasculature maturation. Overall, our study provides valuable knowledge on the properties of MSCs as vasculature-supporting cells and highlights the importance of choosing the application-specific stromal cell source for vascularized organotypic models.

show abstract

Section: Methodsmentioning

confidence: 99%

Vasculogenic Potency of Bone Marrow- and Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells Results in Differing Vascular Network Phenotypes in a Microfluidic Chip

Mykuliak

Yrjänäinen

Mäki

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…In one study, Honkamaki et al developed a layered 3D construct by embedding electrospun poly(L,D-lactide) fibers in collagen hydrogel. The layer-by-layer scaffold shows better resemblance to the native microenvironment, thereby guiding the growth and neurite development of iPSC-derived neuronal cells [ 250 ]. In another recent study, the authors used a 3D hydrogel structure as a base collector instead of a common metal collector to develop a unique hydrogel-assisted electrospinning process.…”

Section: Types Of Hydrogel Units Platforms and Fabrication Technologiesmentioning

confidence: 99%

Engineering Hydrogels for the Development of Three-Dimensional In Vitro Models

Maji

Lee

2022

IJMS

View full text Add to dashboard Cite

The superiority of in vitro 3D cultures over conventional 2D cell cultures is well recognized by the scientific community for its relevance in mimicking the native tissue architecture and functionality. The recent paradigm shift in the field of tissue engineering toward the development of 3D in vitro models can be realized with its myriad of applications, including drug screening, developing alternative diagnostics, and regenerative medicine. Hydrogels are considered the most suitable biomaterial for developing an in vitro model owing to their similarity in features to the extracellular microenvironment of native tissue. In this review article, recent progress in the use of hydrogel-based biomaterial for the development of 3D in vitro biomimetic tissue models is highlighted. Discussions of hydrogel sources and the latest hybrid system with different combinations of biopolymers are also presented. The hydrogel crosslinking mechanism and design consideration are summarized, followed by different types of available hydrogel module systems along with recent microfabrication technologies. We also present the latest developments in engineering hydrogel-based 3D in vitro models targeting specific tissues. Finally, we discuss the challenges surrounding current in vitro platforms and 3D models in the light of future perspectives for an improved biomimetic in vitro organ system.

show abstract

“…In addition to the tested commercial software, the open-source software ImageJ contains tracing ( Lee et al, 1994 ) and analysis ( Arganda-Carreras et al, 2010 ) tools that could be used for analysing interconnected networks. In addition, ImageJ can be used to analyse neuronal directionality and orientation in a robust manner and is free to use ( Honkamäki et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Practical guide for preparation, computational reconstruction and analysis of 3D human neuronal networks in control and ischaemic conditions

et al. 2022

View full text Add to dashboard Cite

To obtain commensurate numerical data of neuronal network morphology in vitro, network analysis needs to follow consistent guidelines. Important factors in successful analysis are sample uniformity, suitability of the analysis method for extracting relevant data and the use of established metrics. However, for the analysis of 3D neuronal cultures, there is little coherence in the analysis methods and metrics used in different studies. Here, we present a framework for the analysis of neuronal networks in 3D. First, we selected a hydrogel that supported the growth of human pluripotent stem cell-derived cortical neurons. Second, we tested and compared two software programs for tracing multi-neuron images in three dimensions and optimized a workflow for neuronal analysis using software that was considered highly suitable for this purpose. Third, as a proof of concept, we exposed 3D neuronal networks to oxygen-glucose deprivation- and ionomycin-induced damage and showed morphological differences between the damaged networks and control samples utilizing the proposed analysis workflow. With the optimized workflow, we present a protocol for preparing, challenging, imaging and analysing 3D human neuronal cultures.

show abstract

Novel method to produce a layered 3D scaffold for human pluripotent stem cell-derived neuronal cells

Cited by 11 publications

References 50 publications

Vasculogenic Potency of Bone Marrow- and Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells Results in Differing Vascular Network Phenotypes in a Microfluidic Chip

Vasculogenic Potency of Bone Marrow- and Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells Results in Differing Vascular Network Phenotypes in a Microfluidic Chip

Engineering Hydrogels for the Development of Three-Dimensional In Vitro Models

Practical guide for preparation, computational reconstruction and analysis of 3D human neuronal networks in control and ischaemic conditions

Contact Info

Product

Resources

About