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细胞功能测定

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MTT Cell Proliferation Assay

MTT 5-dimethylthiazol-2-yl)-25-diphenyltetrazolium bromide assay first described by Mosmann in 1983 is based on the ability of a mitochondrial dehydrogenase enzyme from viable cells to cleave the tetr ...

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NK-cell cytotoxicity assay

Outline:To measure NK cell killing suitable target cells are labeled with 51Cr washed and incubated together with the killer cells (and treatments). Large amounts of label is released into the superna ...

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H-Thymidine Uptake by Cultured Cells

H-Thymidine Uptake by Cultured CellsLEVEL II Materials Fibroblast cells in log phase growthCa Mg free-phosphate buffered saline (PBSA)5% (w/v) Glutaraldehyde (GTA)2% (w/v) Perchloric Acid (PCA)Subbed ...

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cell proliferation assay

cell proliferation assaybefore start:thaw cells from liquid nitrogen grow in 75cc flask (T75) in Fischer's medium MM (maintenance medium) until content gets cloudy and starts to turn yellow (usually w ...

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Quality Control Considerations

IntroductionQuality is important in all aspects of tissue culture since the quality of materials used i.e. media and other reagents) will affect the quality of the cultures and products derived from ...

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Preparation of low density, collagenase-digested splenocytes Protocol

Preparation of low density collagenase-digested splenocytes Protocol 1. Dilute 2 ml of 4000 U/ml Collagenase D as follows: 1 ml into 9 ml HBSS/Ca++/Mg++ (=400U/ml) and 1 ml into 39 ml HBSS/Ca++/Mg++ ...

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Defrosting Eukaryotic Cells Frozen In Liquid N2

Defrosting Eukaryotic Cells Frozen In Liquid N2 Reagents / Solutions Frozen cell line. Growth media (e.g. RPMI1640) Foetal calf serum (FCS). ProtocolMake 20ml growth media/10% FCS for each vial of ce ...

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Eukaryote Growth Dynamics

Eukaryote Growth DynamicsLEVEL II Materials Suspension cultures set up from Exercise 12.8Sterile transfer pipettesMaterials for viability counting (Exercise 12.7) Procedure After 12 hours aseptically ...

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Use of the Hemacytometer for the Determination of Cell Numbers

Use of the Hemacytometer for the Determination of Cell Numbers Counting cells by the use of a hemacytometer is a convenient and practical method of determining cell numbers in the case that the Coulte ...

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Viability Cell Count

Viability Cell CountMaterials Suspension culture of cellsSterile transfer pipettesStock 0.2% (w/v) Trypan blueHemacytometer and microscope Procedure Gently swirl a suspension culture to distribute the ...

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Hemacytometer Reference Guide

Load both sides of the hemacytometer with the cell suspension - covering both counting grids. Count the number of cells in 5 out of 9 squares from each counting grid (count a total of 10 squares). Div ...

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Recommended working medium, trypsin volume and cell inoculation density

Recommended working medium trypsin volume and cell inoculation densityAuthor: Long-Cheng LiSource: Abstract: Useful data for cell subculture with different culture vessels. ...

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Transfer of Eukaryote Suspension Cultures

Transfer of Eukaryote Suspension CulturesLEVEL II Materials Fibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Dis ...

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Culture of endometrial explants

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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Hemacytometer Workbook

This workbook was developed for use with Module 2 of the InVitro Insights Cell Culture Training Program developed by Becton Dickinson. The problem that follow are presented in increasing degress of co ...

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Infection with retroviruses

Infection with retroviruses It is well established with avian retroviruses that cells are most efficiently infected just after they are trypsinized. Trypsinization does two things. First it apparently ...

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Establishment of a Primary Culture

Establishment of a Primary CultureLEVEL III Materials Chick embryo (approximately 8 days old)70% (v/v) ethanol for swabbingSterile scissors forceps and probesSterile petri platesPhosphate buffered sal ...

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Staining protocols for placental explant cultures

Staining protocols for placental explant culturesWhole mount staining.The protocol given uses a single primary antibody with nuclear counterstaining but it can be extended to double antibody staining. ...

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Direct tissue explant method for derivation and study of extravillous trophoblast

From early post-implantation stages to approximately 18 weeks' gestation cytotrophoblast at the tips of anchoring villi at the placental periphery proliferate to form columns. From the distal edges of ...

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Components and Supplementation of Various Culture Media

Components and Supplementation of Various Culture MediaAbbreviations: DMEM = Dulbecco's Modified Eagle's Medium FCS = Fetal Calf (or Bovine) Serum GPS = Glutamine-Penicillin-Streptomycin BME = b-Merca ...

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