细胞技术

In general terms cultured cells require a sterile environment and a supply of nutrients for growth. In addition the culture environment should be stable in terms of pH and temperature. Over the last ...

CULTURE MEDIUM Ingredients these catalog numbers refer to products from Sigma; products from other suppliers can also be used Powdered Hams F-12 (N6760) Powdered MEM D-valine modification (M7395) Feta ...

Preparation of Embryonic ExtractMaterials1. Ice on tray. 2. Dissection tools: 1 pair of large scissors; 2 pairs of fine scissors; 1 pair of coarse forceps; 2 pairs of fine forceps; soak in 95% EtOH. 3 ...

Culture of Endometrial Explants and Peri-implantation Conceptuses to Monitor Synthesis and Secretion of Proteins and Prostaglandins P. J. Hansen and J. G. Betts Dairy Science Dept. University of Flori ...

Cryopreserved Cells (Single donor) ...

MN Analysis : Blood Lymphocytes MN in Human Lymphocytes (method description)Lymphocyte isolationLymphocytes were isolated using Ficoll-Paque density gradients. Blood was diluted 1:1 with phosphat ...

ES Cell Culture and ManipulationCulturing ES cellsMedia High glucose DMEM (-pyruvate -glutamine)20% Heat-inactivated Fetal calf serum (can vary by cell type be sure!!)1X l-glutamine1X Penicillin/stre ...

Analysis of ES Cell Clones by Mini-SouthernInitial Cloning 1. Aspirate selective media off the plates containing colonies of interest. Wash the plates twice with PBS. After aspirating the second was ...

EMBRYONIC STEM CELLS PROTOCOL If you are embarking in growing ES cells be prepared to refeed them DAILY. All procedures should be carried out using sterile techniques. The growth and maintenance ...

b-Galactosidase Color Assay for Cultured Cells 1. For transient expression assays wait until 2 days after the transfection to begin the color assay. Aspirate the media and rinse the cells twice with ...

Electroporation of ES cellsCells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for 1-2 electroporations. P ...

ES and TS cell freezing/thawingNeeded:ES cell freezing medium (2x)2x ES cell freezing medium should be made up fresh each time it is to be used and should comprise freshly prepared 60% DMEM+ 20% FCS a ...

Basic Method for Indirect Immunofluorescence Labeling BackgroundThis is the method for indirect immunofluorescence labeling; that is the antibodies do not have the fluorescent dye attached. Indirect l ...

Differentiate ES cells into cardiac myocytesDay -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.____________________Day 1: Trypsinized t ...

Differentiate ES cells into cystic embryoid bodiesDay -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.______________Day 1: Trypsinized t ...

Preparation of RNA from Cultured CellsNotes: When preparing solutions for use with RNA use only MilliQ water and disposable sterile Tissue Culture plasticware. Also if preparing working solutions fro ...

Preparing 48-Well Plate FeedersIn preparing 48-well plates a dilution must be made from the regulardensity of 3.5 x 105 cells/ml. The required density for 48-well plates is: 8.2 x 104 cells/ml ( ...

TYPES OF CELLS GROWN IN CULTURETissue culture is often a generic term that refers to both organ culture and cell culture and the terms are often used interchangeably. Cell cultures are derived from ei ...

Preparation of Feeder Layers And SNL Stocks1.1. Gelatinizing PlatesPrepare plates by covering surface with 0.1% Gelatin solution: Plate Size (cm) Amount of Gelatin (ml) 3 2 6 4 10 9 15 18Swirl the pla ...

Preparation of NeoR Murine Embryonic FibroblastsThree weeks prior to embryonic fibroblast isolation a PGK-neo male mouse is mated to a heterozygote female. 13.5 days to 14.5 days after the plug is obs ...