Gastruloids as in vitro models of embryonic blood development with spatial and temporal resolution

Gastruloids are three-dimensional embryonic organoids that reproduce key features of early mammalian development in vitro with unique scalability, accessibility, and spatiotemporal similarity to real embryos. Recently, we adapted gastruloid culture conditions to promote cardiovascular development. In this work, we extended these conditions to capture features of embryonic blood development through a combination of immunophenotyping, detailed transcriptomics analysis, and identification of blood stem/progenitor cell potency. We uncovered the emergence of blood progenitor and erythroid-like cell populations in late gastruloids and showed the multipotent clonogenic capacity of these cells, both in vitro and after transplantation into irradiated mice. We also identified the spatial localization near a vessel-like plexus in the anterior of gastruloids with similarities to the emergence of blood stem cells in the embryo. These results highlight the potential and applicability of gastruloids to the in vitro study of complex processes in embryonic blood development with spatiotemporal fidelity.

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Development in a Dish—In Vitro Models of Mammalian Embryonic Development

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.655993 ◽

2021 ◽

Vol 9 ◽

Author(s):

Yasmine el Azhar ◽

Katharina F. Sonnen

Keyword(s):

Embryonic Development ◽

Molecular Mechanisms ◽

Three Dimensional ◽

In Vitro Models ◽

Model Systems ◽

Model Organisms ◽

Mammalian Development ◽

Complex Processes ◽

Mechanisms Of Development

Despite decades of research, the complex processes of embryonic development are not fully understood. The study of mammalian development poses particular challenges such as low numbers of embryos, difficulties in culturing embryos in vitro, and the time to generate mutant lines. With new approaches we can now address questions that had to remain unanswered in the past. One big contribution to studying the molecular mechanisms of development are two- and three-dimensional in vitro model systems derived from pluripotent stem cells. These models, such as blastoids, gastruloids, and organoids, enable high-throughput screens and straightforward gene editing for functional testing without the need to generate mutant model organisms. Furthermore, their use reduces the number of animals needed for research and allows the study of human development. Here, we outline and discuss recent advances in such in vitro model systems to investigate pre-implantation and post-implantation development.

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Decellularized Skeletal Muscles Support the Generation of In Vitro Neuromuscular Tissue Models

Applied Sciences ◽

10.3390/app11209485 ◽

2021 ◽

Vol 11 (20) ◽

pp. 9485

Author(s):

Paolo Raffa ◽

Maria Easler ◽

Francesca Cecchinato ◽

Beatrice Auletta ◽

Valentina Scattolini ◽

...

Keyword(s):

Skeletal Muscle ◽

Three Dimensional ◽

Culture Conditions ◽

In Vitro Models ◽

Easy Access ◽

3D In Vitro Model ◽

Multiple Cell ◽

Vitro Model

Decellularized skeletal muscle (dSkM) constructs have received much attention in recent years due to the versatility of their applications in vitro. In search of adequate in vitro models of the skeletal muscle tissue, the dSkM offers great advantages in terms of the preservation of native-tissue complexity, including three-dimensional organization, the presence of residual signaling molecules within the construct, and their myogenic and neurotrophic abilities. Here, we attempted to develop a 3D model of neuromuscular tissue. To do so, we repopulated rat dSkM with human primary myogenic cells along with murine fibroblasts and we coupled them with organotypic rat spinal cord samples. Such culture conditions not only maintained multiple cell type viability in a long-term experimental setup, but also resulted in functionally active construct capable of contraction. In addition, we have developed a customized culture system which enabled easy access, imaging, and analysis of in vitro engineered co-cultures. This work demonstrates the ability of dSkM to support the development of a contractile 3D in vitro model of neuromuscular tissue fit for long-term experimental evaluations.

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3D Culture Modelling: An Emerging Approach for Translational Cancer Research in Sarcomas

Current Medicinal Chemistry ◽

10.2174/0929867326666191212162102 ◽

2020 ◽

Vol 27 (29) ◽

pp. 4778-4788 ◽

Cited By ~ 1

Author(s):

Victoria Heredia-Soto ◽

Andrés Redondo ◽

José Juan Pozo Kreilinger ◽

Virginia Martínez-Marín ◽

Alberto Berjón ◽

...

Keyword(s):

High Throughput Screening ◽

Cancer Biology ◽

Soft Tissues ◽

Three Dimensional ◽

3D Culture ◽

Resistance Mechanisms ◽

In Vitro Models ◽

Tumour Spheroids ◽

Current Therapies

Sarcomas are tumours of mesenchymal origin, which can arise in bone or soft tissues. They are rare but frequently quite aggressive and with a poor outcome. New approaches are needed to characterise these tumours and their resistance mechanisms to current therapies, responsible for tumour recurrence and treatment failure. This review is focused on the potential of three-dimensional (3D) in vitro models, including multicellular tumour spheroids (MCTS) and organoids, and the latest data about their utility for the study on important properties for tumour development. The use of spheroids as a particularly valuable alternative for compound high throughput screening (HTS) in different areas of cancer biology is also discussed, which enables the identification of new therapeutic opportunities in commonly resistant tumours.

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Organoids of the female reproductive tract

Journal of Molecular Medicine ◽

10.1007/s00109-020-02028-0 ◽

2021 ◽

Vol 99 (4) ◽

pp. 531-553 ◽

Cited By ~ 1

Author(s):

Cindrilla Chumduri ◽

Margherita Y. Turco

Keyword(s):

Reproductive Tract ◽

Personalised Medicine ◽

Three Dimensional ◽

In Vitro Models ◽

Experimental Models ◽

Female Reproductive Tract ◽

Cellular Heterogeneity ◽

Dynamic Regulation ◽

Pregnancy And Childbirth

AbstractHealthy functioning of the female reproductive tract (FRT) depends on balanced and dynamic regulation by hormones during the menstrual cycle, pregnancy and childbirth. The mucosal epithelial lining of different regions of the FRT—ovaries, fallopian tubes, uterus, cervix and vagina—facilitates the selective transport of gametes and successful transfer of the zygote to the uterus where it implants and pregnancy takes place. It also prevents pathogen entry. Recent developments in three-dimensional (3D) organoid systems from the FRT now provide crucial experimental models that recapitulate the cellular heterogeneity and physiological, anatomical and functional properties of the organ in vitro. In this review, we summarise the state of the art on organoids generated from different regions of the FRT. We discuss the potential applications of these powerful in vitro models to study normal physiology, fertility, infections, diseases, drug discovery and personalised medicine.

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Mimicking Tumor Hypoxia in Non-Small Cell Lung Cancer Employing Three-Dimensional In Vitro Models

Cells ◽

10.3390/cells10010141 ◽

2021 ◽

Vol 10 (1) ◽

pp. 141

Author(s):

Iwona Ziółkowska-Suchanek

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Cell Lung Cancer ◽

Tumor Hypoxia ◽

Three Dimensional ◽

In Vitro Models ◽

Small Cell ◽

Small Cell Lung ◽

Multicellular Tumor Spheroid

Hypoxia is the most common microenvironment feature of lung cancer tumors, which affects cancer progression, metastasis and metabolism. Oxygen induces both proteomic and genomic changes within tumor cells, which cause many alternations in the tumor microenvironment (TME). This review defines current knowledge in the field of tumor hypoxia in non-small cell lung cancer (NSCLC), including biology, biomarkers, in vitro and in vivo studies and also hypoxia imaging and detection. While classic two-dimensional (2D) in vitro research models reveal some hypoxia dependent manifestations, three-dimensional (3D) cell culture models more accurately replicate the hypoxic TME. In this study, a systematic review of the current NSCLC 3D models that have been able to mimic the hypoxic TME is presented. The multicellular tumor spheroid, organoids, scaffolds, microfluidic devices and 3D bioprinting currently being utilized in NSCLC hypoxia studies are reviewed. Additionally, the utilization of 3D in vitro models for exploring biological and therapeutic parameters in the future is described.

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