ProtocolforCellFusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, they should be centrifuged at a 64.4 xg on the IEC clinical centrifuge (in 50 ml tubes) for two minutes. The media should be removed and the cells resuspended in fresh media. This ensures that only the healthiest cells will fall to the bottom of the tube. Centrifugation at......閱讀全文
Growing－cells
No two cell lines behave exactly the same, so you must learn the peculiarities, or personality, of each of the cell lines with which you work. Irresp
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell Lines June 10, 1990 Rosalie Veile Purpose: To grow lymphoblastoid cells for permanent stora
Plastic－dishes－for－growing－cells
There are two kinds of dishes used to grow tissue culture cells. Those that are designed for adherent cells have been treated chemically to promote
Freezing－and－Thawing－cells
Freezing and Thawing cellsFreezingIt is best to freeze cells that are growing rapidly. With adherent cells, it is easiest to set up 100 mm dishes, giv
Production－of－neuronpreferential－lentiviral－vectors
實驗概要Adenoviral vectors widely used to transfer foreign genes into neuronal cells possess tropism for glial cells and are toxic to infected cells.
Protocol－for－Cell－Fusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium:? 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
Growing－feederindependent－embryonic－stem－cells§
We use feeder-independent ES cell lines derived from the 129/Ola strain of mice (Nichols et al., Development 110, p.1341, 1990). These cells are easy
The－OP9DL1－System:－Generation－of－TLymphocytes－from－Embryonic4
TROUBLESHOOTINGProblem:?The OP9 cells are more than 80%-90% confluent.Solution:?It is important when creating working stocks of OP9?cells for freezing
Freezing－and－Thawing－of－Mammalian－Cell－Lines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.
Live－imaging－with－Drosophila－tissue－culture－cells1
Introduction Live imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows fol
細胞培養常規操作
常規操作(主要內容如下)·?????????Aseptic Technique·?????????Culture Vessels·?????????Cell Counting·?????????Primary Culture·?????????Maintenance of Cell Line?·??
CellTrace?－CFSE－Cell－Proliferation－Kit
實驗概要The CellTrace? ?CFSE Cell Proliferation Kit provides a versatile and well-retained ?cell-tracing reagent in a convenient and easy-to-use form. The
Cryopreservation－of－cell－cultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial con
ES－Cell－Culture－and－Manipulation3
Care and Handling of Feeder Layer Cells STO/SNL cells are a derivative of the standard STO cell line. They are transformed with a LIF producing pl
Subculture－of－Adherent－Cell－Lines
AimAdherent cell lines will grow in vitro until they have covered the surface area available or the medium is depleted of nutrients. At this point the
Subculture－of－Suspension－Cell－Lines
Aim In general terms cultures derived from blood (e.g. lymphocytes) grow in suspension. Cells may grow as single cells or in clumps (e.g. EBV transfo
Growing－Cells－in－Geltrex?－Reduced－Growth－Factor－Basement－Membrane－Matrix
實驗概要Basement ?membranes are continuous sheets of specialized extracellular matrix ?that form an interface between endothelial, epithelial, muscle, or
Creation－and－Use－of－Infectious－Virus－Vector
Creation and use of your infectious vector:Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 ml of media. (This can be scaled up if desired).The f
Thawing－Cells－(Schreibers－protocol)
Thaw vial quickly in 37癈 water. Caution - vial can explode. Transfer cells to sterile, 15 mL centrifuge tube. Add?50 祃?warm?FBS?(fetal bovine
QUALITATIVE－ANALYSIS－OF－DNA－FRAGMENTATION－BY－AGAROSE－GEL－ELECTROPHORESIS
1. IntroductionNuclear morphology changes characteristic of apoptosis appear within the cell together with a distinctive biochemical event: the endonu
CREATION－AND－USE－OF－YOUR－INFECTIOUS－VECTOR
實驗概要CREATION AND USE OF YOUR INFECTIOUS VECTOR實驗步驟Day 1? ? ? ? 1. Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 mL of media. (This can be scal
Routine－Culturing－of－ES－Cells
Cell are normally passaged every 2-3 days, this is important to avoid differentiation.Signs of differentiation are:-i) colonies are surrounded by flat
TOP10－chemically－competent－cells
OverviewThis protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the B
ES－Cell－Culture－and－Manipulation2
Picking ES cell clonesOne or two days before picking colonies prepare 24-well plates of feeders. You can also use alternate protocols that utilize 96-
慢病毒轉染肝細胞方法
Lentivirus Transduction of Hematopoietic CellsMing-Jie Li and John J. RossiDivision of Molecular Biology, Beckman Research Institute of the City of Ho
Method:－Lymphocyte－Transformation
Method: Lymphocyte Transformation May 30, 1990 Rosalie Veile Principle: Lymphocytes are transformed to establish cell lines. Mononuclear
Dual－and－TripleCo...－（二）
主要試劑 1. Blocking reagent after coating: 10% fetal calf serum (FCS) in phosphate-buffered saline (PBS), pH 7.4. 2. Cells suspended in complete cultu
FIXATION－and－DNA－Staining－for－Cell－Cycle－Analysis
Background This method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to e
The－OP9DL1－System:－Generation－of－TLymphocytes－from－Embryonic1
The OP9-DL1 System: Generation of T-Lymphocytes from Embryonic or Hematopoietic Stem Cells In VitroRoxanne Holmes?and?Juan Carlos Zú?iga-Pflücker1Sunn