Organotypicraftcultureofprimaryhumankeratinocytes(PHKs)
Organotypic raft culture of primary human keratinocytes (PHKs)NotesUse 106 keratinocytes per raft.Typical ProtocolTrypsinize the PHKs off the flask and resuspend in (say) 2ml raft medium.See separate protocol for raft mediumRemove the medium from the collagen plugs and overlay with the new medium containing PHKs.Incubate for 1-2 days to allow the PHKs to settle and become confluent.Autoclave the f......阅读全文
Organotypic－raft－culture－of－primary－human－keratinocytes－(PHKs)
Organotypic raft culture of primary human keratinocytes (PHKs)NotesUse 106 keratinocytes per raft.Typical ProtocolTrypsinize the PHKs off the flask an
Ex－Vivo－Human－Primary－Mesenchymal－Stem－Cells－(MSCs)－Culture
Human bone marrow contains mesenchymal progenitors (mesenchymal stem cells, MSCs). MSCs produce adventitial cells in the human bone marrow microenvir
Primary－brain－cell－isolation－and－culture
1. Cerebella were removed from 7-day-old mice and passed through Nitex nylon netting (80 μm pore size) into primary cell system containing 20% (v/
Exercise－12.10－－Establishment－of－a－Primary－Culture
Exercise 12.10 - Establishment of a Primary CultureLEVEL IIIMaterialsChick embryo (approximately 8 days old)70% (v/v) ethanol for swabbingSterile scis
Raft－culture－media－(aka－Green’s－media)
Raft culture media (aka Green’s media)3 parts DMEM (including glutamine or glutamax)1 part Ham’s F125% FCSvarious supplements (not everybody uses the
A－primary－cell－culture－model－of－rabbit－uroepithelium
Isolation of Epithelial Cells from Rabbit Bladders 1. Animal experiments were performed in accordance with the Animal Use and Care Committee. 2. Urina
Primary－type－II－pneumonocyte－Isolation－and－culture
Pulmonary alveolar type II cells carry out highly specialized functions that include the synthesis, secretion, and reutilization of surfactant, a
Isolation－of－human－primary－airway－smooth－muscle
1. Primary cultures of human primary airway smooth muscle (ASM) cells were prepared from explants of airway smooth muscle. 2. The human trachea tiss
Human－B－cell－isolation－and－culture
实验概要This protocol provides a general protocol for human B cell isolation and culture.实验步骤B cell isolation1. Donor blood was obtained with informed con
Human－B－cell－isolation－and－culture
B cell isolation1. Donor blood was obtained with informed consent under guidelines issued by the Medical Ethics Committee. 2. Mononuclear cell fractio
Primary－Culture－of－Pulmonary－Arterial－Smooth－Muscle－Cells
Primary Culture of Pulmonary Arterial Smooth Muscle Cells 1.Primary cultures of Pulmonary Arterial Smooth Muscle Cells (PASMCs) were isolated from hum
Isolation－of－human－primary－gastric－mucosa－epithelial－cells
1. A piece of gastric mucosa (∼3 cm2) was obtained from the normal appearing mucosa of the stomach at surgery. 2. The patients were proven to be no H.
Isolation－of－human－primary－ovarian－surface－epithelial－cells
1. Surface epithelial cells from normal ovaries from surgical residual specimen were isolated using standard and Institutional Review (References)
Culture－of－human－venous－smooth－muscle－cells
Culture of human venous smooth muscle cells 1. Explants of the 2 groups of skins (control subjects and patients with varicose veins) were prepared
Culture－of－Human－Embryonic－Stem－Cells－(hESC)
All cell lines are initially grown according to the supplier's protocols but we are adapting them to one simple protocol outlined below:6-well pla
Isolation－and－Culture－of－Human－Liver－Stem－Cells
Materials and Methods1. Human hepatocytes were isolated from fresh surgical specimens of patients undergoing hepatectomies. Healthy liver tissue (
Isolation－and－Culture－of－human－vascular－smooth－muscle－cells
1. Human vascular smooth muscle cells (SMCs) were isolated from human saphenous vein.2. Small moistened pieces of tissue (6 mm2) were placed intima
Isolation－and－Culture－of－Human－Brain－Tumor－Stem－Cell
The isolation, culture, identification, and purification of stem cells from primary human brain tumors of different phenotypes have marked capacit
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, t
Collagen－gel－containing－3T3－fibroblasts－dermal－equivalent－for－raft－culture
Collagen gel containing 3T3 fibroblasts (dermal equivalent for raft culture)Ingredients for 6 x collagen matrices in a 6-well plate;Roughly 3x106 J2-3
Isolation－and－culture－of－human－pulmonary－artery－smooth－muscle－cells－...
Isolation and culture of human pulmonary artery smooth muscle cells (PASMCs)1. The human pulmonary arteries were opened to expose the endothelial su
Culture－of－human－renal－proximal－tubule－cells－and－renal－cortical－fibroblasts
1. The method for primary culture of human renal proximal tubule cells (PTC) and renal cortical fibroblasts (CF) is described in detail.2. Renal cor
Isolation－and－Culture－of－Multipotent－Stem－Cells－from－Human－Bone－Morrow
Bone marrow contains three types of stem cells:1. Hematopoietic stem cells give rise to the three classes of blood cells that are found in the circu
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
Trabecular－cell－monolayer－culture
Originally described in 1979 and more recently modified by Stamer et al primary trabecular monolayer cell culture has been a cornerstone for inves
Mouse－keratinocyte－cultures
PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al., Methods Enzymol
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
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
Isolation－of－human－uterine－smooth－muscle－cells
1. Approval to utilize human tissue was given by Hospital's Committee on Research Involving Human Subjects. 2. Human myometrial and leiomyoma tiss
Culture－of－retinal－endothelial－cells－and－pericytes
Isolation of retinal microvessels1. Eyes were obtained and transported on ice.2. Eyes were cut circumferentially 3 mm posterior to the limbus, the v