KARYOTYPINGESCELLS
An actively growing culture of cells is required, i e 2 - 3 d ES cell culture. The total number of cells needs to be between 106 - 107 cells.N B Read notes at end of method.1) Harvest cells in the usual manner, into a conical bottom centrifuge tube.2) Centrifuge to remove medium.3) Resuspend cell button by flicking the bottom of the tube.4) Add 8 ml of warmed KC1 solution to each tube and mix gently with a pipette.5)......阅读全文
KARYOTYPING－ES－CELLS
An actively growing culture of cells is required, i e 2 - 3 d ES cell culture. The total number of cells needs to be between 106 - 107 cells.N B Read
Electroporation－of－ES－cells
Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for
Differentiate－ES－cells－into－glial－cells－and－neurons
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.___________________Day 1: Trypsiniz
FACS－Analysis－of－ES－Cells
Isolate cells and dissociate to single cell suspension (can use Gibco Cell Dissociation Buffer, Accutase or Trypsin)Wash with 10% FBS/DMEM:F12For surf
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
Screen－ES－cells－by－Southern－Blot
Digest DNA in 96-well plateTo each well add:4ul 10Xbuffer4ul Enzyme0.4ul Spermidine(0.4M)31.6ul H2O37‡C 19h, then add 4ul loading dye to each well. Lo
Differentiate－ES－cells－into－cardiac－myocytes
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.____________________Day 1: Trypsini
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
Differentiate－ES－cells－into－cystic－embryoid－bodies
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.______________Day 1: Trypsinized th
Human－Embryonic－Stem－(ES)－Cell－Protocols——Splitting－Human－ES－cells－on－MEFs
based on splitting onto one plateWarm collagenase IV split media to 37 °C in a water bath.Aspirate media off of cell culture plate.Add the following a
Human－Embryonic－Stem－(ES)－Cell－Protocols—Splitting－Human－ES－cells－onMatrige
based on splitting onto on plateWarm collagenase media to 37°C in a water bath.Aspirate media off of cell culture plate.Add the following amount of co
ELECTROPORATION－OF－ES－CELLS－AND－ISOLATION－OF－H/R－CLONES
Need 1.5-2 x 107 cells from a 2 day culture.1. Cells are harvested as normal, washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in cold
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 (
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
Differentiating－Neural－Stem－Cells－into－Neurons－and－Glial－Cells
实验概要The protocols in this section describe the steps involved in differentiating neural stem cells (NSC) to neurons, astrocytes, and oligodendrocyte
Trypsinizing－cells
There are many procedures with which to trypsinize cells. All include washing the cell monolayer with TD, or in rare cases, with VE. This removes seru
Lyophilizing－Cells
Inoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized.Incubate the culture at 37°C with vigorous shak
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.
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. Irrespe
Lyophilizing－Cells
Lyophilizing CellsInoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized.Incubate the culture at 37°C
Human－Embryonic－Stem－(ES)－Cell－Protocols——General－notes－on－ES－cell－culture
hES media has a two week shelf life.hES cells should be cultured in 4, 6, 24, 48, 96 well plates. Growing cells in flasks is not recommended because i
ES细胞分化培养实验——培养皿ES细胞集落
实验材料ES 细胞单一胚胎样体仪器、耗材细菌培养皿ES 分化培养基实验步骤1. 准备下列试剂和材料：悬滴培养的 ES 细胞单一胚胎样体100 mm 细菌培养皿ES 分化培养基2. 在 100 mm 细菌培养皿中加 10 ml 预热的分化培养基。3. 从温箱中取出培养 2 天的悬滴培养物。小心翻转培养
Counting－ES－cell－Chromosomes
(original reference: "tissue culture made easy" by Christian LaMantia from the Magnuson lab)1) Plate cells onto gelatinized plates (35 or 60 mm) witho
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
Preparation－Of－Peripheral－Blood－Cells－For－Chromosome－Analysis
实验概要Lymphocytes are differentiated cells which normally do not undergo subsequent cell divisions. By culturing lymphocytes in the presence of a mito
Preparing－cells－and...
实验概要The method provides a protocol and tips for BrdU staining in tissue sections.Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic
Subculturing－Adherent－Cells
实验概要The following protocol describes a general procedure for subculturing adherent mammalian cells in culture.主要试剂1. Complete growth medium, pre-warme
Isolation－of－papillary－cells
实验概要This protocols provides a general protocol for isolation of papillary cells.实验步骤Isolation of renal papillary cells1. For isolation of papillary c
Fluorescent－Staining－of－Cells
1. Fluorescent phalloidin in methanol. Phallacidin does not work as well. Dilute 10 ul 330 nM stock into 500 ul PBS for each large coverslip.2. PB