Rotary Cell Culture System

The rotary cell culture system is a device designed to grow 3D cells clusters (tissue, cancer tumors and virus cultures) in microgravity (outside the body). The advantage here is to obtain larger and 3D cell cultures with structural and chemical characteristics similar to normal tissue, in comparison with lab-grown cell cultures.

It allows to test chemotherapy agents on a patient’s own cancer cells ex vivo.

2D-LEC cultures

wound scratch assayIn these assays, (Lymphatic) Endothelial Cells ((L)ECs) are seeded as monolayers on culture plates or onto the surface of matrix-coated plates.

Such 2D cultures can be used to evaluate the effects of putative lymphangiogenic stimulators or inhibitors on specific LEC properties. While none of the 2D cultures can undergo all steps of lymphatic vessel formation, these systems contribute to analyze cell activities (e.g. gene expression profiling), cell proliferation, apoptosis, adhesion, migration (wound scratch assay, boyden chamber assay) and morphogenesis (tubulogenesis).

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

In this assay, Lymphatic Endothelial Cells (LECs) monolayer is scratched out. LECs migrate to wound the scar.

This assay allows the assessment of LEC migratory capabilities. Mitomycin C can be added in the culture medium to inhibit cellular proliferation.

Plates are observed under a phase contrast microscope and the percentage of area recovered by migrating cells is determined.

Boyden chambers are a useful tool to study cell migration and cell invasion such as chemotaxis, haptotaxis and transmigration. It consists of a cylindrical cell culture insert nested inside the well of a cell culture plate. The insert contains a polycarbonate membrane at the bottom with a defined pore size.

Cells are seeded in the top of the insert in serum-free media, while serum or similar chemoattractant are placed in the well below. Migratory cells move through the pores toward the chemoattractant below and can be stained or quantified in a plate reader.

Invasive cells may be similarly measured by the placement of a coating of extracellular matrix proteins on top of the membrane.

3D-LEC Cultures

3D cultures bridge the gap between in vitro and in vivo assays and are available to follow the sprouting process and Lymphatic Endothelial Cells (LEC) morphogenesis. The lymphatic-ring assay appears as a potent tool for the study of pathological lymphangiogenesis, defined as the abnormal formation of new lymphatic vessels from preexisting ones.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

tubulogenesisLEC are seeded on matrix-coated substrates and organized into capillary-like structures.

Results are expressed as the area density of tube-like structures, defined as the area occupied by tubes divided by the total area of the studied field.

sproutingThe spheroid sprouting assay consists of a self-aggregation of LEC embedded in a 3D matrix composed of type I collagen. LEC originating from the spheroids progressively invade the gel and migrate either individually as unicellular sprouts (single cells) or collectively as complex capillary-like structures. The area occupied by the migrated cells and the spatial distribution of cells around the initial cell aggregate are determined through an original computerized method.

High-grade glioma-initiating cells derived from normal astrocytes

The transformed astrocytes are obtained from normal mouse astrocytes undergoing gamma irradiation after TGF-alpha treatment.

In this in vitro model, transformation causes major losses of astrocyte-specific proteins and functions and the acquisition of metabolic adaptations that favor intermediate metabolites production for increased macromolecule biosynthesis.

The cells display all canonical features of cancerous transformation, immortalization, uncontrolled growth, cytogenetic alterations, and give rise to high-grade gliomas when injected into nude mice. In culture, they constitute a homogenous population of transformed astrocytes particularly suited for quantitative biochemical comparison with their normal counterparts, represented by primary cultures of astrocytes. It could similarly be applied to the evaluation of the effects of treatments aimed at correcting the consequences of cell transformation.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

Clonogenic assay

Clonogenic assay or colony formation assay is an in vitro cell survival assay based on the ability of a single cell to grow into a colony. The colony is defined to consist of at least 50 cells.

This assay essentially tests every cell in the population for its ability to undergo “unlimited” division.

Clonogenic assay is the method of choice to determine cell reproductive death after treatment with ionizing radiation, but can also be used to determine the effectiveness of other cytotoxic agents.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

Chondrocytic dedifferentiation model

Although dedifferentiation is a phenomenon described for decades, molecular events underlying the process are not yet fully elucidated. As a consequence, dedifferentiation still constitutes a major brake in vitro for chondrocytes expansion, thereby preventing cartilage regeneration using expanded autologous chondrocytes.

To provide better understanding of chondrocytes biology, the researchers at GIGA applied human osteoarthritis hip chondrocytes cultured in monolayer in a short-term dedifferentiation model (during 14 days). In this model, molecular events as well as substances able to delay or reverse dedifferentiation process can be tested.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

Mesenchymal differentiation model

The researchers from GIGA have a large expertise in the isolation, expansion and differentiation of mesenchymal synovial fibroblasts (SVF) isolated from synovial tissue.

SVF monolayer cultures represent an interesting model to study adipogenic, chondrogenic and osteogenic properties.

The isolated SVF:

  • Are plastic adherent cells with an important proliferation potential
  • Express mesenchymal stem cell immunophenotype (CD105, CD73 and CD90 and lack of CD45 expression)
  • Are able to differentiate, in vitro in adipocytes, osteocytes and chondrocytes under specific conditions

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

Osteoblast-like cells cultures in monolayer

In this model, osteoblast-like cells lines are cultured in monolayer. It allows a better understanding of signaling pathways in various experimental conditions or after exposure to various agents.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

Tumor model on CAM

In this model, glioblastoma cells are implanted on chicken chorioallantoic membrane (CAM) at day 11 after fertilization. This implantation results in tumor growth showing an histology similar to glioblastoma multiform in patients, with diffuse pleiomorphic infiltrate of fibrillar and stellate cells, neoangiogenesis, edema, and areas of necrosis.

The growth of tumors on CAM provides a more rapid, low cost, and ethically sustainable alternative.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

Organotypic culture models

Organotypic culture involves the combination of cells in a specific ratio to create a component of an organ. Researchers at GIGA have expertise in development of organotypic cultures of normal and tumor epithelial cells, involving the growing cells in a 3D environment.

They developed organotypic culture systems that permit epithelial cells to proliferate and differentiate at an air – liquid interface on a dermal equivalent support. Normal keratinocytes stratify and fully differentiate in a manner similar to the normal squamous epithelial tissues.

This culture system is more similar biochemically and physiologically to in vivo tissue. Human epidermis reconstructed in vitro may be used as the best alternative for the in vitro testing of the toxicology and efficiency of products for topical use, as well as in the treatment of skin burns and chronic skin ulcers.

The developed models include:

The dermal equivalent is an in vitro model of the dermal layer of skin. It is constructed by seeding dermal fibroblasts into a collagen gel. Other cell types may be incorporated into the dermal equivalent to increase the complexity of the model.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.

Human keratinocytes and melanocytes cultured in vitro are grown on a biological matrix (dead de-epidermized human dermis) and the system is kept at an air-liquid interface, in a suitable culturing medium, until a stratified human epidermis is formed, maintaining the histological characteristics of the epidermis in vivo. De-epidermized dermis model is a closer approximation to the in vivo situation of formation and maintenance of the mature epidermis.

Contact [info@b2h.be] us to discuss how these capabilities can forward your projects! We will help you develop tailored solutions.