Cryopreservationofcellcultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial contamination and should be subconfluent (confluent cultures may be less amenable to freezing).2. Remove the growth medium and wash twice in PBS (B0.1 ml cm2).3. Add prewarmed (37 1C) trypsin solution (0.5 ml per 25 cm2) to cover the cell monolayer, recap the flask and incu......阅读全文
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
Placental－trophoblast－and－chorionic－cell－cultures
1. Placental trophoblast and chorionic trophoblast cells were prepared using a modification of the method. 2. Term human placentae and chorion tissue
Rodent－Retinal－Ganglion－Cell－Cultures
实验概要Central neurons lose the ability for axonal regrowth during development and typically do not regenerate their axons following axotomy once the
Mitochondrial－DNA－Isolation－from－Somatic－Embryogenic－Cell－Cultures－of－Larix
Mitochondrial DNA is isolated by a modification of the methods described by Wilson and Chourey (1984) and Radetzky (1990). Cell cultures at four d
Preserving－yeast－cultures
Short term storage Yeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags. Medium term stora
ORGANOTYPIC－KIDNEY－CULTURES
-embryos are dissected from timed-pregnant mice from 11.5 d.p.c. to 13.5 d.p.c.-metanephroi and associated ureteric buds are microdissected and placed
Hippocampal－Neuron－Cultures
实验概要The protocol provides a method of hippocampal neuron cultures.主要试剂Begin by timing the pregnant mouse at E17-E19 days of gestation. Have ready the
ORGANOTYPIC－LIMB－CULTURES
-embryos are dissected from timed-pregnant mice from 10.5 - 11.5 d.p.c.-limb buds are microdissected and placed in holding medium (L15 medium suppleme
Growing－Overnight－Cultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-ca
Mouse－keratinocyte－cultures
PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al., Methods Enzymol
Transfer－of－Eukaryote－Suspension－Cultures
Materials Fibroblast suspension culture Tissue culture laminar flow hood Media appropriate to culture line used Disposable pipettes
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
实验概要 AMNIOTIC FLUID CULTURES ON COVERSLIPS 主要试剂 Solutions: Colcemid working solution: 10 mcg/ml Colcemid in Hank's Balanced Salt Solution
Primary－Cultures－fo...
实验概要The following protocol provides a method of primary cultures for IHC – viability assays.实验步骤1. Preparation of primary mesencephalic cultures 1)
Dissociated－Cultures－of－Cerebellar－Neurons
Dissociated Cultures of Cerebellar NeuronsHank Dudek (617-355-4735)Protocolisolate cerebella (ÒCbÓ)cut off head into plate with HHGNhold nose with lar
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
I. Purpose:Amniotic fluid may be used for prenatal diagnosis of aneuploidy or other structural abnormalities. II. Culture Procedure:A. Aseptic techniq
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose: A: Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspec
SOLID－TUMOR－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose: A. Samples of solid tumors or lymph nodes may be sent from patients with cancer. These samples should be processed directly and also set
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
实验概要 Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. susp
Shellless－cultures－of－chick－embryos
This experiment allows you to observe the development of a chick embryo outside its shell. Cultured chicks are also more accessible for manipulation.
Barretts－esophageal－epithelial－and－fibroblast－primary－cultures
1. Biopsy specimens for tissue culture were immediately placed on ice in primary cell culture system. 2. Within 4 hours from the time of the biopsy,
Method:－Removal－of－Yeast－Contamination－from－Lymphoblast－Cultures
Purpose:This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when s
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell Lines June 10, 1990 Rosalie Veile Purpose: To grow lymphoblastoid cells for permanent stora
A－quick－RNA－miniprep－for－Neurospora－mycelial－cultures
Most RNA isolation techniques currently in use have been developed for the processing of large quantities of material. These typically involve multip
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 ·
Primary－cultures－of－intrahepatic－bile－duct－epithelial－cells－isolated－and...
Primary cultures of intrahepatic bile duct epithelial cells isolated and cultured1. Liver was surgically removed and perfused via the hepatic vein.2.
Isolation－of－normal－mammary－epithelial－cells
1. A small piece of tissue from each specimen was removed and minced. 2. The tissue was digested with collagenase overnight. 3. To remove the collagen
Isolation－of－human－prostatic－epithelial－cells
1. A small piece of tissue from each specimen was removed and minced. 2. The tissue was digested with collagenase overnight. 3. To remove the collagen
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
Activation－and－Expansion－of－Human－Treg－Cells
实验概要This protocol is intended for activation and expansion of human Treg cells isolated with the Dynal® CD4 CD25 Treg Kit (Cat. no. 113.23D). The exp
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