FreezingandThawingofMammalianCellLines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.FreezingPreparationCells are to be frozen in liquid nitrogen, so make sure your canisters are relatively full of nitrogen and you have room. Cells should be healthy (>90% viability) and growing in log phase. You will also require sterile 1 mL cryo-vials; they have a scr......阅读全文
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.Fre
Cell－Thawing/Cell－Freezing－Protocol
Freezing Cells:Cells should be growing well or known to be in log phaseCount, collect and pellet cells in a 15mL test tubeResuspend in freezing media
Cell－Thawing/Cell－Freezing－Protocol
Freezing Cells:Cells should be growing well or known to be in log phaseCount, collect and pellet cells in a 15mL test tubeResuspend in freezing media
ES－and－TS－cell－freezing/thawing
Needed: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
ES－and－TS－cell－freezing/thawing
实验概要ES and TS cell freezing/thawing.主要试剂ES cell freezing medium (2x) 2x ES cell freezing medium should be made up fresh each time it is to be
细胞培养——细胞保藏
Working Cell Bank (Contributed by Nanci Donacki)Provides detailed protocol for establishing a working cell bank Master Cell Bank (Contributed by Na
Freezing－and－Thawing－of－MEFs
Author: Shalini Jain and Hariom YadavAffiliation: Animal Biochemistry Division, National Dairy Research Institute, Karnal-132001, Haryana, IndiaDate A
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
Cryopreservation－of－Cell－Lines
AimThe protocol below describes the use of passive methods involving an electric -80ºC freezer for the cryopreservation of cell cultures. ECACC routin
Method:－Preparation－of－Lymphoblastoid－Cell－Lines－for－Long－Term－Storage
Method: Preparation of Lymphoblastoid Cell Lines for Long Term StorageMay 30, 1990Rosalie VeilePurpose:To store cell lines in a form that will insure
Resuscitation－of－Frozen－Cell－Lines
AimMany cultures obtained from a culture collection, such as ECACC, will arrive frozen and in order to use them the cells must be thawed and put into
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
AimIn general terms cultures derived from blood (e.g. lymphocytes) grow in suspension. Cells may grow as single cells or in clumps (e.g. EBV transform
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell LinesJune 10, 1990Rosalie VeilePurpose:To grow lymphoblastoid cells for permanent storage and for DNA extracti
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
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
Subculture－of－SemiAdherent－Cell－Lines
AimSome cultures grow as a mixed population (e.g. B95-8 - marmoset) where a proportion of cells do not attach to the tissue culture flask and remain i
Tissue－Culture－Methods1
I. 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 us
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
细胞培养常规操作
常规操作(主要内容如下)· Aseptic Technique· Culture Vessels· Cell Counting· Primary Culture· Maintenance of Cell Line ·
Method:－Lymphoblastoid－Cell－Lines－from－Frozen－Whole－Blood
Method: Lymphoblastoid Cell Lines from Frozen Whole BloodMay 31, 1990Rosalie VeilePurpose:Blood Samples can be stored frozen as a backup in case an LC
Tissue－Culture－Methods3
REFERENCES:R. Ian Freshney, Culture of Animal cells: A manual of basic techniques, Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach student should m
ES－Cell－Culture－and－Manipulation
MediaHigh glucose DMEM (-pyruvate, -glutamine)20% Heat-inactivated Fetal calf serum (can vary by cell type, be sure!!)1X l-glutamine1X Penicillin/stre
Human－Embryonic－Stem－(ES)－Cell－Protocols——Freezing－Human－ES－Cells
Collagenase cells for approximately 7 minutes at 37 °C (until edges of colonies are curling up).With a 5 ml pipet, gently pipet and scrape colonies f
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates
AbstractThe kinetic characteristics and regulation of aspartate carbamoyltransferase activity were studied in lysates and cell extracts of Helicobacte
胚胎干细胞培养
Media and Solution required for ES Cell Culture (Bowtell Lab) Routine Culturing of ES Cells (Bowtell Lab) Routine Splitting and freezing of cells (
Human－Embryonic－Stem－(ES)－Cell－Protocols——Thawing－Human－ES－cells
Remove Human ES cells from liquid nitrogen storage tank. Fill out a freeze/thaw form.Thaw cryovial by gently swirling in waterbath until only a small
Preparation－of－cytoplasmic－extracts－forthe－application－in－acellfree－system
DescriptionCells are grown to 80% confluency, then harvested, washed and disrupted in KPM buffer by freezing-thawing cycles with liquid nitrogen essen
Preparation－of－cytoplasmic－extracts－for－the－application－inacellfree－system
Characteristics of this procedure:Cells are grown to 80% confluency, then harvested, washed and disrupted in KPM buffer by freezing-thawing cycles wit