Spheroid Transfection
Spheroid Transfection - Related Videos
07:40
Gene Transfection toward Spheroid Cells on Micropatterned Culture Plates for Genetically-modified Cell Transplantation
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Cited by 9 •
2015
This protocol describes a cell transplantation system using genetically modified, injectable spheroids. Cell spheroids are cultured on micropatterned culture plates and recovered after gene introduction using polyplex nanomicelles. This system facilitates prolonged transgene expression from the transplanted cells in host animals while maintaining the innate function of the cells.
07:28
An Introduction to Transfection
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2023
Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery, whereas insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for transfected RNA is the cytoplasm, DNA must be transported to the nucleus for effective transfection. There, the...
03:20
Splitting Spheroids for Sub-culturing and Shipping: A Procedure for Propagating and Transferring Spheroids from Small Bowel Neuroendocrine Tumor
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2023
This video demonstrates a strategy to split and propagate spheroids developed from small bowel neuroendocrine tumor cells. These spheroids have small bowel neuroendocrine tumor markers and can be propagated and further used to test anti-cancer drugs.
04:29
Immunofluorescent Staining of Spheroids: A Technique to Analyze Spheroids for Expression of Intra- and Extracellular Antigen Expression
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2023
This video describes the technique of immunofluorescent staining of spheroids to understand the differentiation of the fibroblasts or CAFs. This protocol helps detect expression of intra- and extracellular antigens, including deposition of ECM proteins within the spheroids.
02:59
Embedding 3D Spheroids in Agarose Gel Drops: A Technique to Preserve Spheroids for Downstream Analysis
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2023
This video describes the technique to embed 3D spheroids in agarose gel drops for preservation. The embedded 3D spheroids can be further used for downstream analysis.
FAQs
Delivery is constrained by the spheroid’s compact architecture. Dense cell layers and extracellular matrix can limit how far nucleic acids and delivery systems move through the aggregate. As a result, exposure at the outside does not necessarily produce uniform access to internal cells, making penetration a central determinant of the outcome.
Both reagent-based and physical delivery methods must overcome the same structural barriers: dense cell layers and extracellular matrix within the spheroid. Their relevance depends on whether they can promote access beyond the outer region while preserving cell viability. The delivery approach therefore affects both how broadly nucleic acids reach the aggregate and how reliably the model remains usable.
Spheroid transfection can reveal effects that are not apparent in two-dimensional culture because cells retain three-dimensional cell-cell interactions, extracellular matrix context, and tissue-like gradients. Consequently, changes in gene expression or behavior can be examined within a more complex multicellular structure, helping determine whether a response depends on spatial organization rather than a flat culture context.
A basic workflow must account for the nucleic acid cargo, the spheroid’s three-dimensional structure, and the selected reagent-based or physical delivery approach. The aggregate must be exposed to a method capable of reaching cells beyond the surface, while viability is preserved. The key procedural concern is therefore effective access without disrupting the tissue-like model.
Applications include developmental biology, cancer research, drug-response studies, tissue engineering, and gene-function analysis. In these settings, researchers can connect altered gene expression with coordinated behavior in a three-dimensional cell model. This is particularly useful when responses may depend on cell-cell interactions, extracellular matrix, or gradients that are not represented fully in two-dimensional cultures.
Assessment should consider both whether nucleic acids reach cells across the spheroid and whether the aggregate remains viable. Delivery limited to outer layers may not represent manipulation throughout the structure, while reduced viability can complicate interpretation. Considering access and viability together helps researchers decide whether observed responses reflect gene alteration or damage to the biological model.