Cell Culture Basics

The process of removing cells from an animal or plant and growing them in a controlled environment.

A cell culture that is directly taken from a living organism and grown in a lab setting.

A cell culture that has undergone at least one division and can continue dividing indefinitely.

The process of transferring cells from one culture vessel to another with a fresh growth medium.

Cells that have a limited lifespan and eventually stop dividing.

Cells that have been genetically modified to allow them to divide indefinitely.

Cells taken directly from living tissue and cultured in a lab.

Cells derived from cancerous tissues or genetically modified to divide indefinitely.

A type of cell culture media that lacks phenol red, a pH indicator dye. This media is often used for colorimetric assays or fluorescence experiments where the color of phenol red would interfere with the readouts.

A common component in many cell cultures, derived from blood. It's a complex mixture of nutrients, growth factors, and other components essential for cell growth and survival.

A process where serum is heated to 56°C for 30 minutes. This inactivates the complement system (a part of the immune response) and eliminates potential mycoplasma contamination.

Essential nutrients like amino acids, vitamins, inorganic minerals, fats, and nucleic acid derivatives that are crucial for cell growth and metabolism.

Proteins like fibronectin and laminin present in serum that help cells adhere to surfaces in the culture environment.

Growth factors like FGF, EGF, and PDGF present in serum that stimulate cell division and proliferation.

Proteins like albumin and transferrin that bind and transport important molecules like vitamins, fats, hormones, and iron within the cell culture environment.

Antibiotics are frequently used in cell cultures to prevent contamination by bacteria or fungi. This is especially crucial when aseptic techniques are employed.

Stem cells from a newborn's umbilical cord blood can be stored for later use in medical treatments, such as transplants for diseases like leukemia or sickle cell anemia.

Stem cell transplantation can help people with certain cancers like leukemia and lymphoma, bone marrow diseases, anemia like sickle cell disease, and some immune disorders.

Cord blood is being researched for its potential use in treating other serious conditions like Parkinson's disease and diabetes.

Cryopreservation is a method to freeze and store cells at extremely low temperatures (-200°C) so they can be used later.

Cryoprotectants like DMSO or glycerol prevent ice crystal formation during freezing, protecting cells from damage.

Cryovials are labeled with important information like date, cell type, and passage number to keep track of the frozen cells.

Before freezing, cells are washed with Phosphate Buffered Saline (PBS) and trypsin.

A freezing medium containing a cryoprotectant is added to the cells before freezing.

A modified version of Eagle's MEM containing approximately four times the vitamins, amino acids, and two to four times the glucose. DMEM is commonly used for cells with faster growth rates and difficult attachment, like epithelial and fibroblast cells. It also includes iron and phenol red.

A complex medium typically used for human lymphocytes, but can support a wide range of cell types. It is a modified version of DMEM.

A modification of DMEM that is well-suited for difficult proliferating, low-density cell cultures. It is useful for hybrid cell selection after fusion and selection of transformed cells following DNA transfection.

A modification of Basal Medium 5A that supports the growth of primary cultures derived from diverse tissues. It is a general-purpose medium for both primary and established cell lines.

A versatile medium used for the serum-free growth of Chinese Hamster Ovary (CHO) cells, as well as other mammalian cells. It is a general-purpose medium.

Contamination in cell culture comes in two main forms: chemical and biological. Chemical contaminants include impurities in media, sera, and water, as well as endotoxins, plasticizers, and detergents. Biological contaminants include bacteria, molds, yeasts, viruses, mycoplasma, and cross-contamination from other cell lines.

Many bacterial strains can grow in the commonly used media for mammalian cell culture. Bacterial contamination can severely affect cell growth and experimental results.

Aseptic technique is a set of procedures used to prevent microbial contamination. It involves practices such as sterilizing equipment, working in a sterile environment, and minimizing the time materials are exposed to the air.

Mesenchymal stem cells (MSCs) are adult stem cells with a high capacity for self-renewal and differentiation, capable of becoming various cell types.

MSCs can differentiate into various cell types, including bone cells (osteoblasts), cartilage cells (chondrocytes), and fat cells (adipocytes).

Osteoblasts, chondrocytes, adipocytes.

MSCs have the potential to differentiate into muscle cells (myocytes), nerve cells (neurocytes), and stromal cells.

MSCs can be isolated from various sources such as bone marrow, adipose tissue, umbilical cord, peripheral blood, placental tissue, synovial fluid, and dental pulp.

MSCs have been studied for various therapeutic applications, including tissue repair, regenerative medicine, and cell-based therapies for various diseases.

The process of removing cells from a living organism and cultivating them in a controlled environment.

The process of separating bone marrow from a mouse, using a combination of anesthesia, dissection, and sterile techniques.

A technique used to analyze and sort individual cells based on their physical and biological properties. It uses lasers and fluorescent dyes to identify and separate cells based on their size, granularity, and specific protein expression.

The process of using fluorescently-labeled antibodies to bind to specific proteins or molecules on the surface or inside of cells, making them visible under a flow cytometer.

A method of identifying apoptotic cells by measuring the exposure of phosphatidylserine (PS) on the cell surface using fluorescently labeled Annexin V. PS is normally located on the inner leaflet of the cell membrane but flips to the outer membrane during apoptosis.

A dye that can penetrate cells only when their plasma membrane is compromised, indicating cell death. It's often used in combination with Annexin V to differentiate early and late stage apoptosis.

Apoptosis is a programmed cell death that occurs in a highly regulated process, involves DNA fragmentation and cell shrinkage.

Necrosis is a form of cell death that occurs when the cell is damaged by external factors, often leading to inflammation and tissue damage. It involves cell swelling, loss of membrane integrity, and release of cellular contents.

Flow cytometry uses the principle of scattering light from a laser beam to identify and quantify different cell populations. This scattering patterns give information about the cell's size and granularity.

Flow cytometer has various applications, including: 1) Immunophenotyping: identifying cell types based on their surface markers. 2) Cell cycle analysis: determining the stage of the cell cycle (e.g., G0, G1, S, G2, M). 3) Cell viability analysis: measuring the number of live and dead cells in a sample. 4) Cell sorting: isolating specific cell populations based on their characteristics.