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细胞功能测定

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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 J2-3T3s (a fully confluent T75?) 1.5mL 10x reconstitu ...

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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 protocol was adapted from one prepared by Aydin Guzelog ...

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Culture of Endometrial Explants and Peri-implantation Conceptuses to Monitor Synthesis and Secretion of Proteins and Prostaglandins

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 ...

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Method: Maintaining Lymphoblastoid Cell Lines

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 ...

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MN in Human Lymphocytes (method description)

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 ...

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Organotypic raft culture of primary human keratinocytes (PHKs)

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 ...

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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 up for short term culture of tumor cells. Care must ...

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TISSUE CULTURE ON COVERSLIPS

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 ...

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Tissue Culture of PtK1 cells

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 ...

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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. suspected mosaicism confirmation of a new chromosome disorde ...

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Transfer of Eukaryote Suspension Cultures

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 ...

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Growth, Maintenance and Transfection of Suspension Adapted 293-EBNA cells

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 ...

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Aseptic Technique and Good Cell Culture Practice

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 ...

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Resuscitation of Frozen Cell Lines

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 ...

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Subculture of Adherent Cell Lines

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 ...

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Subculture of Semi-Adherent Cell Lines

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 ...

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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 routinely use a programmable rate controlled freezer (Pl ...

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Testing for Bacteria and Fungi

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 ...

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Detection of Mycoplasma by Culture

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 ...

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Testing for Mycoplasma by Indirect DNA Stain (Hoechst 33258 stain)

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 ...

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