Non-enzymatic Tissue Culture

Non-enzymatic tissue culture is the in vitro growth and maintenance of tissue fragments without using digestive enzymes to separate their cells, preserving the native organization of the starting material. Under aseptic conditions, tissue is placed on a nutrient medium that supplies water, mineral salts, sugars, and, when needed, plant growth regulators or other signaling factors; cells then survive, divide, and reorganize within the cultured tissue. This approach supports studies of tissue development, regeneration, cell interactions, and responses to environmental or chemical stimuli. By retaining tissue architecture, non-enzymatic culture can complement dissociated-cell systems and provide biologically relevant models for experimental biology.

Non-enzymatic Tissue Culture - Related Videos

Research

JoVE Journal - Biology

Non-enzymatic, Serum-free Tissue Culture of Pre-invasive Breast Lesions for Spontaneous Generation of Mammospheres

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2014

Primary cell culture using intact tissue organoids provides a model system that mimics the multi-cellular in vivo microenvironment. We developed a serum-free primary breast epithelium tissue culture model that perpetuates mixed cell culture lineages and exhibits differentiated morphology, without enzymatic tissue disruption. Breast organoids remain viable for >6 months.

Education

JoVE Science Education - Engineering

Histotypic Tissue Culture

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2023

Although two-dimensional tissue culture has been common for some time, cells behave more realistically in a three-dimensional culture, and more closely mimics native tissue. This video introduces histotypic tissue culture, where the growth and propagation of one cell line is done in an engineered three-dimensional matrix to reach high cell density. Here, we show the harvesting of cells from donor tissue, followed by cell culture on an engineered construct.

Whole Organ Tissue Culture

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2023

Whole organs can be cultured ex vivo using specialized bioreactors with the goal of repairing or replacing entire organs. This method uses a donor organ that is stripped of all cells, leaving behind the three-dimensional structure, which is then repopulated with new cells. This video demonstrates the whole organ culture of lungs and shows how a dynamic culture that mimics the mechanical stimulation in the body is needed to induce native tissue properties.

Plant Tissue Culture

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2020

Plant tissue culture is widely used in both primary and applied science. Applications range from plant development studies to functional gene studies, crop improvement, commercial micropropagation, virus elimination, and conservation of rare species. Plant tissue culture depends on the ability of plant tissue to give rise to an entire new plant when provided with a growth medium and appropriate environment. This ability of plant cells or tissues is termed ‘totipotency.’ The fundamental steps...

A Simple and Rapid Protocol to Non-enzymatically Dissociate Fresh Human Tissues for the Analysis of Infiltrating Lymphocytes

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Cited by 29 •

2014

This protocol describes the rapid non-enzymatic dissociation of fresh human tissue fragments for qualitative and quantitative assessment of CD45+ cells (lymphocytes/leukocytes) present in various normal and malignant human tissues. Additionally, the supernatant obtained from the primary tissue homogenate can be collected and stored for further analysis or experimentation.

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