Cell Biology & Tissue Culture: Culture Environment, Lab Procedures, and Ethics

Flashcards covering mammalian cell culture environment requirements, specific cell lines, media components, physical parameters like pH and osmolarity, and ethical considerations in biotechnology.

Mammalian Cell Culture Aim

To provide an environment that mimics, to the greatest extent possible, the in vivo environment of a specific cell type.

HeLa

A human cell line derived from ovarian cancer.

DU145

A human cell line derived from prostate cancer.

MCF-7

A human cell line derived from breast cancer.

Vero

An African green monkey Chlorocebus kidney epithelial cell line initiated in 1962.

CHO

A Chinese hamster ovary cell line.

MC3T3

A mouse embryonic calvarium cell line.

MDCK

Madin-Darby canine kidney epithelial cell line.

Culture Temperature

Maintained by an incubator typically at $37^\text{o}C$ to simulate in vivo conditions.

Phenol Red

A pH indicator where yellow/orange indicates acidic (exhausted media or contamination), orange/pink indicates neutral (perfect for growth), and dark pink/red indicates basic (gas imbalance).

Buffering

The process of maintaining a neutral pH despite cellular waste and $CO_2$ changes, often maintained by HEPES.

Viscosity of DMEM with $10\%$ FBS

Measured at $0.94\,cP$, this property influences the movement and diffusion of molecules required for metabolic processes.

DMEM

Dulbecco′s Modified Eagle′s Medium; a basal, fully defined media.

Riboflavin

A vitamin essential to the formation of coenzymes FMN and FAD, involved in energy metabolism and cell respiration.

FBS (Fetal Bovine Serum)

A media supplement derived from cow fetuses that contains essential growth factors but is expensive and batch-dependent.

Growth Factors

Unique cell signaling molecules like FGF (Fibroblast Growth Factor) and EGF (Epidermal Growth Factor) that play roles in cell proliferation and development.

Serum-free media (SFM)

Media that allows specific cell growth in the absence of serum, offering easier purification and more consistent performance but requiring time for cell adaptation.

HEPES

A chemical buffering agent that maintains physiological pH and enzyme structure despite changes in carbon dioxide concentration.

Glutamine

An unstable amino acid supplement needed in cell culture to support growth and function.

PBS (Phosphate buffered saline)

An isotonic salt solution (typically containing $NaCl$, $KCl$, $Na_2HPO_{4}$, and potassium phosphate) that maintains a stable pH around $7.4$.

Osmotic shock

The rupture of cells in a solution caused by a sudden reduction in osmotic pressure.

$5\%\,CO_2$ Purpose

Provides a buffer for closed growing conditions to ensure normal physiological pH is maintained via bicarbonate buffering.

Substrate

A material providing a surface for adherent cells to attach and grow on, mimicking their natural body environment.

Adherent cultures

Cells that require a surface to attach to and grow on, forming a monolayer.

Suspension cultures

Cells that grow freely floating in the culture medium.

Certificate of Analysis

A document that reports and certifies the test results of a product.

Morphology table

A lab record used to capture cell details, media observations, and high/low power microscopic observations.

Informed Consent

A critical ethical requirement prior to tissue collection, historically violated in cases like the HeLa cell line and the Tuskegee Syphilis trial.