FreezingandThawingcells
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, give them fresh medium the day before you freeze them, and freeze them just as they become confluent. If you are working with suspension cells, make sure that they are growing vigorously before you freeze them. 1. Trypsinize the cells if necessary, and then spin ......阅读全文
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
Freezing－and－Thawing－of－MEFs
Author: Shalini Jain and Hariom YadavAffiliation: Animal Biochemistry Division, National Dairy Research Institute, Karnal-132001, Haryana, IndiaDate A
Freezing－Cells
1) Keep prepared solutions on ice.2) Determine total cell count of cells to be frozen. (e.g. 1 X 108 )3) Determine number of vials to be frozen. (e.g.
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
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
Freezing－cells－in－liquid－nitrogen
Take off MediaTrypsinate with 1ml x2 Dulbecco A trypsinAdd 7ml MediaPipette up and down to distribute cells throughout media (i.e. not clumped togethe
Routine－Splitting－and－freezing－of－cells
1. Grow cells to subconfluence in a flask.2. Harvest as per normal and count.3. Spin down 5min 1.2K in benchtop. Resuspend at 1.0 X 106/ml in 10% DMSO
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 serum, hea
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
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
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－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
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
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 (
stem－cell－culture－protocol
实验概要stem cell culture protocol主要试剂cell culture supplies and reagentssEnvironment: cell culture requires a sterile environment, so it needs a separat
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:－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
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
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
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
病毒冷冻保藏技术
实验概要Snap freezing, or flash freezing, is the process by which samples are lowered to temperatures below -70°C very rapidly using dry ice or liquid
Preserving－yeast－cultures
Short term storageYeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags.Medium term storageYeast
Growth,－Maintenance－and－Transfection－of－Suspension－Adapted－293EBNA－cells
ProcedureI. INTRODUCTIONThe 293 EBNA cell line is established from primary embryonal human kidney cells transformed with sheared human adenovirus type
Comparison－of－Enzymatic－and－NonEnzymatic－Means2
MTT Assay on Reattached CellsMSC were seeded in 12-well cell culture dishes with 5.0 × 104 cells per well (≈4.8 cm2). After 5 to 6 days of culture, co
Culture－of－BEND－Cells－(Bovine－Endometrial－Cells)
Culture of BEND Cells (Bovine Endometrial Cells)Charles E. Krininger, III and Peter J. Hansen Dept. of Animal Sciences, University of FloridaThis prot
Serum－Thawing－－Heat－Inactivation
Serum Thawing & Heat Inactivation(Chris Cohick from JRH Biosciences catalogue)How to thaw serum:Serum that is stored at -10º C to -40º C is stable for
Thawing－and－Plating－Cryopreserved－Hepatocytes
实验概要This protocol covers thawing and prep of cryopreserved hepatocytes for applications such as metabolic stability, intrinsic clearance, enzyme in