Which trypsin to use

This website uses cookies to ensure you get the best experience on your website. Learn more. Trypsin-EDTA 0. Enzymatic cell dissociation reagent. Catalog Add to Cart. Skip to the end of the images gallery.

Skip to the beginning of the images gallery. Related Products. Scientific Resources. Product Applications. Data and Publications. This is a gentler version than the standard concentration 0. Related Products Related Products. Product Applications This product is designed for use in the following research area s as part of the highlighted workflow stage s.

Research Area. Tissue Dissociation. Transfection and Selection. Expansion and Characterization. Workflow Stages for Neuronal Research. Cell Culture. View publication. Different cell lines should never be processed at the same time. All working areas should be thoroughly cleaned between the preparations of different cell types. Precautions Wear personal protective equipment.

Use proper aseptic technique. Skip to main content Skip to primary sidebar Skip to footer Purpose The following protocol is suggested for the basic instruction of culturing adherent cell lines. A confluent culture is when all the cells are in contact and the entire surface of the culture vessel is covered. Epithelial-like — Cells that are polygonal in shape with more regular dimension, and grow attached to a substrate in discrete patches.

Fibroblastic or fibroblast-like — Cells grown on a substrate that appear elongated and bipolar, frequently forming swirls in heavy cultures Hybridomas — Cell line formed by fusing two different but related cells. Immortal or Continuous cell line — Cells that have been adapted to grow indefinite in culture.

Lymphoblast-like — Cells that are spherical in shape and are usually grown in suspension without attaching to a surface. Monolayer culture system — Cells that primarily grow attached to glass or treated plastic in a uniform layer.

Passage number — The number of sub-cultures the cells have undergone. Passage number should be recorded with each sub-culture. Primary culture — Cells that are derived from and organism and placed into a suitable culture environment.

Senescence — cumulative changes to molecular and cellular structure that disrupts metabolism with the passage of time, resulting in deterioration and eventually leading to cell death. Subculture or passaging — removal of medium and transfer of cells from a previous culture into fresh growth medium that enables continued propagation of the cell line.

Suspension culture system — Free floating cells in a culture system. Tissue culture — The general term for removal of cells, tissues, or organs from an animal or plant and subsequent placement into an artificial environment conducive to growth. Quiescence — A state of quietness or inactivity, a state of a cell when it is not dividing.

Lab task wipes Disinfectant Lens tissue paper Cell culture media Dissociation media such as Trypsin 0. Procedure Preparation of cell growth medium. Check the information pertaining to the cell line to identify what media type, additives and recommendations should be used.

Follow manufacturer recommendation for working volume of culture vessels. Examination of cells Cells should be checked microscopically often to ensure they are healthy and growing as expected. Attached cells should be mainly attached to the bottom of the flask, round and plump or elongated in shape and refracting light around their membrane. Discard cells if they are detaching in large number and look shriveled and grainy or dark in color, if they do not appear to be growing at all, or appear to have been contaminated.

Split-Ratios Split ratios can be used to ensure cells should be ready for an experiment. Check the guidelines for the cell lines in use. Some slow growing cells may not grow if a high split ratio is used. Some fast growing cells may require a high split ratio to make sure they do not overgrow. Sub-culturing or passaging Remove and discard the spent cell culture media from the culture vessel. Wash cells using a balanced salt solution without calcium and magnesium.

Approximately 2mL per 10cm2 cultured surface area. Gently add wash solution to the side of the vessel opposite the attached cell layer to avoid disturbing the cell layer. Gently rock the vessel back and forth several times to wash the cell layer. Remove and discard the wash solution from the culture vessel.

Repeat the wash step for a total of two washes.