cultures Protocols

Category: Neuroscience: Neuron Cell Culture Protocols

Hippocampal Neuron Culture Protocol, methods. Jean Siao, Ph.D. Tsirka Lab. - [Read Hippocampal Neuron Cell Cultures]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Cultured mammalian cells are used extensively in cell biology studies; it requires a number of special skills in order to be able to preserve the structure, function, behavior and biology of the cells. This unit describes the basic skills required to maintain and preserve cell cultures: aseptic technique, medium characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. - [Read Basic Techniques for Mammalian Cell Tissue Culture Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques

Introduction to Animal Cell Culture. Corning Life Science PDF. Key concepts to Cell Cultures an Introductory Manual with definitions and explanations. - [Read Introduction to Animal Cell Culture - Corning Life Science PDF.]

Category: Bacteria Transformation Protocols

Protocol generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). - [Read Preparation and High-efficiency Transformation of Competent E. coli]

Category: Cell Biology and Cell Culture: Hepatocyte Cell Culture

USC Liver Transplant Surgery Program and Center for Liver Disease. Preparation of primary cultures of rat hepatocytes (or freshly isolated hepatocytes). - [Read Preparation of primary cultures of rat hepatocytes (or freshly isolated hepatocytes)]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

This unit describes how to detect and recognize bacterial, fungal, or mycoplasma contamination and how to verify the presence of mycoplasma. Strategies are described for dealing with culture contamination, if necessary. - [Read Assessing and Controlling Microbial Contamination in Cell Cultures Protocol]

Category: Cell Biology and Cell Culture: Hepatocyte Cell Culture

Primary culture of hepatocytes is an in vitro model widely used to ... protocols for cytochromes P450IIIA in vivo and in primary cultures of animal and ...Human Hepatocyte Culture Protocol - [Read Human Hepatocyte Culture Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

Derivation of Mouse Embryonic Fibroblast Primary Cultures. MEF. MEFP - [Read Derivation of Mouse Embryonic Fibroblast Primary Cultures DOC]

Category: Cell Culture Protocols: Cell Storing and Cell Freezing Protocols

Cryogenic preservation (storage below -100°C) of cell cultures is widely used to maintain backups or reserves of cells without the associated effort and expense of feeding and caring for them. The success of the freezing process depends on four critical areas: Proper handling and gentle harvesting of the cultures; Correct use of the cryoprotective agent; A controlled rate of freezing; Storage under proper cryogenic conditions. - [Read Cryogenic Preservation and Storage of Animal Cells Protocol]

Category: E Coli Protocols: E coli Transformation Protocols

Procedure generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). IMPORTANT All steps in this protocol should be carried out aseptically. - [Read Preparation and Transformation of Competent E. coli Protocol]

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

MEF feeders are prepared weekly to provide a substrate for undifferentiated embryonic stem (ES) cells. Primary MEF cells are thawed, established in culture, treated with mitomycin C to halt their proliferation so they cannot overgrow the ES cultures, and then replated onto dishes convenient for ES cell culture. This protocol can also be used to prepare feeder cells from STO fibroblast cell lines. - [Read Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

No cell culture problem is as universal as that of culture loss due to contamination. All cell culture laboratories and cell culture workers have experienced it. Culture contaminants may be biological or chemical, seen or unseen, destructive or seemingly benign, but in all cases they adversely affect both the use of your cell cultures and the quality of your research. Contamination problems can be divided into three classes: Minor annoyances, Serious problems, Major catastrophes. - [Read Understanding and Managing Cell Culture Contamination Protocol]

Category: Cell Culture Protocols: Sterile Technique Protocols

This unit describes some of the ways that a laboratory can deal with the constant threat of microbial contamination in cell cultures. A protocol on aseptic technique is described first. This catch-all term universally appears in any set of instructions pertaining to procedures in which noncontaminating conditions must be maintained. - [Read Aseptic Technique for Cell Culture Protocol]

Category: Cell Culture Protocols: Sterile Technique Protocols

Cultured mammalian cells are used extensively in cell biology studies; it requires a number of special skills in order to be able to preserve the structure, function, behavior and biology of the cells. This unit describes the basic skills required to maintain and preserve cell cultures: aseptic technique, medium characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. - [Read Basic Techniques for Mammalian Cell Tissue Culture Protocol]

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Protocol for primary cultures of HUVECs. Includes: Buffers and reagents (Fetal calf serum, Phosphate Buffer Saline (PBS), Collagenase, Buffer for conservation and transport of umbilical cords, Culture medium); Cell culture. - [Read Protocol for Primary Cultures of HUVECs]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when slightly shaken and lymphocytes will not cluster together as much as normal. If cultures are suspect, a drop of culture can be streaked on a YPD media plate to check for growth of yeast colonies, or a 5 ml sample can be taken to Barnes Diagnostic Center for identification of yeast strain. - [Read Removal of Yeast Contamination from Lymphoblast Cultures Protocol]

Category: Bacteria Transformation Protocols

Protocol reproducibly generates competent cultures of E. coli that yield 1 x 108 to 3 x 108 transformed colonies/µg of plasmid DNA. The protocol works optimally when the bacterial culture is grown at 18°C. If a suitable incubator is not available, a standard bacterial shaker can be set up in a 4°C cold room and regulated to 18°C. - [Read Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells Protocol]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Growth of Primary Cell Culture and Viral Vector Handling Protocols]

Category: Mouse Protocols: Mouse Cell Culture Protocols

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Isolation and Growth of Primary Cell Cultures from Mouse Protocol]

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Protocol describes a method for storing bacterial cultures containing glycerol which are growing in liquid media. - [Read Storage of Bacterial Cultures Growing in Liquid Media Protocol]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

Protocol describes a method for mycoplasma detection using Hoechst 33258, a fluorescent dye that stains DNA. Staining of particulate DNA matter around the cell nucleus (on the cell surface or in the cytoplasm) is indicative of mycoplasma contamination. - [Read Detection of Mycoplasma in Mammalian Cell Cultures Using Fluorescence Microscopy Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Describes the steps in detail to isolate and expand neural stem cells in the form of neurospheres from tissue dissections of the post-natal mouse brain. Procedures for the long term passage of neurospheres and the cryopreservation of neurospheres are also provided. In addition to the guidelines and tips for generating neurosphere cultures, we describe the method to prepare neurospheres for analysis by light microscopy. - [Read Neural Stem Cell Culture: Neurosphere Generation, Microscopical Analysis and Cryopreservation]

Category: Neuroscience: Neuron Cell Culture Protocols

Preparation of cultures, Ansselin et al., 1998. Acta Chururgica Austriaca 30. - [Read Successfully culturing Schwann cells from adult peripheral nerve]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular.... - [Read Fluorescent In Situ Transcription in Cells and Tissues Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Advantages of Freezing Cell Cultures, Practical Aspects of Cell Freezing, Cryoprotection, Storage Vessels, Labeling and Recordkeeping, Recovery, Managing a Cell Repository, cell selection, Problem Solving Suggestions. Corning. by John A. Ryan - [Read General Guide for Cryogenically Storing Animal Cell Cultures]