Category: Cell Culture

Commonly Used Cell Culture Vessels

Culture vessel types Surface area Culture flask T-25 25 cm2 T-75 75 cm2 T-175 175 cm2 Culture dish 35 mm…

Preparation of Metaphase Chromosome Spreads from Adherent Cell Line

Overview Preparation of metaphase chromosome spread is an important technique in cytogenetics. Metaphase chromosome spread is used for karyotyping including…

Handling and Aliquoting Frozen Stock of Serum

The serum is commercially supplied in different sizes with 500 ml being the most common size. Since repeated freezing and…

Preparing Laminar Flow Hood for Cell Culture Work

Laminar flow hood (Class II) provides a sterile environment for handling cell culture. To make the laminar flow hood ready…

Protocol – Heat Inactivation of Serum

Overview Heat inactivation of serum refers to the process which involves treatment of serum at a higher temperature to inactivate…

Protocol – Feeding Suspension Culture (Complete Replacement of Medium)

Complete replacement of culture medium from suspension culture involves cell harvesting by centrifugation and resuspension of the cell pellet in fresh growth medium. This method is ideally the best way to feed suspension culture and is often used when a. you observe dead or sick cells/suspended particles in culture; b. medium color (phenol red-containing medium) has turned yellow very quickly; c. you can not afford cell loss. Complete replacement of culture medium is ideally the best way to feed suspension culture. Since this procedure takes time and involves an additional centrifugation step to harvest cells, often researchers partially replace the medium that takes just a few minutes. However, partial medium replacement is associated with the loss of cells from the culture.

Protocol – Feeding Suspension Culture (Partial Replacement of Medium)

Replacement of culture medium from suspension culture is a bit tricky job as cells float in the medium. Ideally, cells are harvested from the medium by centrifugation and then resuspended in a fresh culture medium. However, in practice, a part of the culture medium is replaced with a fresh culture medium without harvesting the cells. This procedure is associated with the loss of cells.

Protocol – Feeding Adherent Culture

Feeding adherent culture involves replacing the old consumed medium with the fresh growth medium. The purpose is to supply fresh…

Preparation of Stock Solution of L-Glutamine (200 mM)

Overview L-Glutamine (C5H10N2O3; CAS number: 56-85-9) is an important component of the cell culture medium. It is an alternative energy…

Feeding Cells

Feeding is an essential step to maintain cells in a healthy state in culture. Feeding of a culture is a…

Protocol – Thawing and Revival of Cryopreserved Cells

This protocol describes the revival process of cryopreserved adherent cells. The revival process involves rapid thawing of cryopreserved cells followed by removal of freezing medium which contains cryoprotectant, in this case, DMSO. The freezing medium can be removed immediately just after thawing the cells by centrifugation. Alternatively, cells are allowed to adhere to the culture dish followed by washing and addition of fresh growth medium.

How To Choose Appropriate Cell Line For Research?

Description of basic criteria to select a cell line for your research project.

Protocol – Cryopreservation of Adherent Cells Growing in Serum-free Medium

Overview A serum-free freezing medium is used to preserve cell lines which are maintained in the serum-free growth medium. Serum-free…

Bacterial Contamination in Cell Culture

Bacterial Contamination in Cell Culture

Bacterial contamination is one of the most common cell culture contamination. Poor aseptic culture conditions, including handling, incubator and laminar…