Collagen gel containing 3T3 fibroblasts (dermal equivalent for raft culture)Ingredients for 6 x collagen matrices in a 6-well plate; Roughly 3x106 J2-3T3s (a fully confluent T75?) 1.5mL 10x reconstitu ...
Culture of BEND Cells (Bovine Endometrial Cells) Charles E. Krininger III and Peter J. Hansen Dept. of Animal Sciences University of FloridaThis protocol was adapted from one prepared by Aydin Guzelog ...
Culture of Endometrial Explants and Peri-implantation Conceptuses to Monitor Synthesis and Secretion of Proteins and Prostaglandins P. J. Hansen and J. G. Betts Dairy Science Dept. University of Flori ...
Method: Maintaining Lymphoblastoid Cell LinesJune 10 1990 Rosalie Veile Purpose: To grow lymphoblastoid cells for permanent storage and for DNA extraction. Safety Considerations: All cultured animal a ...
MN in Human Lymphocytes (method description)Lymphocyte isolationLymphocytes were isolated using Ficoll-Paque density gradients. Blood was diluted 1:1 with phosphate buffered saline and layered ont ...
Organotypic raft culture of primary human keratinocytes (PHKs)Notes Use 106 keratinocytes per raft. Typical Protocol Trypsinize the PHKs off the flask and resuspend in (say) 2ml raft medium. See separ ...
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 up for short term culture of tumor cells. Care must ...
I. Purpose:A. Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive e.g. suspected mosaicism confirmation of a new chromosome disorde ...
Please note: lately (2005 - present) we normally culture PtK1 cells in F-12 media but they also grow in other types of media as described below. Mixing F-12 (HAM) Mediundefined ~undefinedSigma stopped making the po ...
I. Purpose:A: Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive e.g. suspected mosaicism confirmation of a new chromosome disorde ...
Materials Fibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Disposable culture flasks Procedure Obtain a culture ...
ProcedureI. INTRODUCTIONThe 293 EBNA cell line is established from primary embryonal human kidney cells transformed with sheared human adenovirus type 5 DNA and Epstein-Barr virus nuclear antigen 1 al ...
AimTo ensure all cell culture procedures are performed to a standard that will prevent contamination from bacteria fungi and mycoplasma and cross contamination with other cell lines.Materials Chloros ...
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 culture. It is vital to thaw cells correctly in orde ...
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 cell lines should be sub-cultured in order to pre ...
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 in suspension. Therefore to maintain this heterogen ...
AimThe protocol below describes the use of passive methods involving an electric -80ºC freezer for the cryopreservation of cell cultures. ECACC routinely use a programmable rate controlled freezer (Pl ...
AimIn cases of gross contamination the naked eye may identify the presence of bacteria and fungi. However it is necessary to detect low-level infections by incubation of cell cultures and/or their pro ...
AimDetection of mycoplasma by culture is the reference method of detection and has a theoretical level of detection of 1 colony-forming unit (cfu). However there are some strains of mycoplasma that ar ...
AimDNA staining methods such as Hoechst staining techniques are quick with results available within 24 hours which compares favorably with 4 weeks for detection by culture. However the staining of cul ...