SubcultureofSuspensionCellLines
AimIn general terms cultures derived from blood (e.g. lymphocytes) grow in suspension. Cells may grow as single cells or in clumps (e.g. EBV transformed lymphoblastoid cell lines). For these types of lines subculture by dilution is relatively easy. But for lines that grow in clumps it may be necessary to bring the cells into a single cell suspension by centrifugation and resuspension by pipetting in a smaller volume ......閱讀全文
Subculture－of－Suspension－Cell－Lines
AimIn general terms cultures derived from blood (e.g. lymphocytes) grow in suspension. Cells may grow as single cells or in clumps (e.g. EBV transform
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
Subculture－of－SemiAdherent－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 i
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
細胞培養常規操作
常規操作(主要內容如下)·?????????Aseptic Technique·?????????Culture Vessels·?????????Cell Counting·?????????Primary Culture·?????????Maintenance of Cell Line?·??
Cell－Suspension－Culture－of－Arabidopsis
實驗概要Cell Suspension Culture of Arabidopsis?主要試劑10% (v/v) Household BleachCallus Induction Medium?? ? ?Gamborg's B5 Basal Medium?? ? ?0.5 g/liter M
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
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell LinesJune 10, 1990Rosalie VeilePurpose:To grow lymphoblastoid cells for permanent storage and for DNA extracti
Freezing－and－Thawing－of－Mammalian－Cell－Lines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.Fre
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
Preparation－of－cytospin－from－single－cell－suspension.
Cell number, speed and trivial details affect cytospin .Basic protocol:Prepare a cell suspension of not more than 0.5 x 106?cells / ml of protein-cont
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Fibroblast－Cell－Systems3
Seeding?After cells are thawed:NOTE:?Do not?dispense the entire contents of the cryovial into one T-25 flask!!Remove the cap, being careful not to tou
Tissue－Culture－Methods3
REFERENCES:R. Ian Freshney,?Culture of Animal cells: A manual of basic techniques,?Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach?student?should m
Method:－Preparation－of－Lymphoblastoid－Cell－Lines－for－Long－Term－Storage
Method: Preparation of Lymphoblastoid Cell Lines for Long Term StorageMay 30, 1990Rosalie VeilePurpose:To store cell lines in a form that will insure
Method:－Lymphoblastoid－Cell－Lines－from－Frozen－Whole－Blood
Method: Lymphoblastoid Cell Lines from Frozen Whole BloodMay 31, 1990Rosalie VeilePurpose:Blood Samples can be stored frozen as a backup in case an LC
Fibroblast－Cell－Systems5
Hemacytometer Reference Figure 15.Determine the Cell Count.a. Calculate the?total?cells counted in the four corner squares.1) If the?total?cell count
Cell－Viability－Assay
Dye exclusiona cell suspension is mixed with trypan blue and examined by low-power microscopyMaterialscellsPBSM3hemocytometer0.4 % trypan blue in PBSm
Freezing－and－Thawing－of－MEFs
Author:?Shalini Jain and Hariom YadavAffiliation:?Animal Biochemistry Division, National Dairy Research Institute, Karnal-132001, Haryana, IndiaDate A
Growing－cells
No two cell lines behave exactly the same, so you must learn the peculiarities, or personality, of each of the cell lines with which you work. Irrespe
CO2恒溫搖床解決人胚腎－293－(HEK293)－細胞結團問題（一）
人胚腎 293 (HEK293)? 細胞在重組蛋白表達中是最常見的宿主細胞。 這類細胞能夠表達大量的膜蛋白，如 G 蛋白偶聯受體? (GPCR) ，是無法在最常見的生物制藥生產宿主，如：中國倉鼠卵巢 (CHO) 細胞中作表達。 HEK293 雖然是蛋白表達的極好宿主，然而 HEK293 細胞
Transfecting－Suspension－Cells
實驗概要將轉移基因整合到細胞染色體DNA上,形成穩定表達轉移基因的細胞系。?實驗原理? ? 細胞轉染技術是目前廣泛應用于病毒基因結構與功能以及基因調控等的研究。細胞轉染可分為短暫轉染和穩定(或永久) 轉染兩種。在短暫轉染中,被轉染基因并不整合至細胞染色體中,因而不能隨細胞分裂而傳代。轉入病毒基因的轉
Selection－of－Transfected－Suspension－Cells
Contributor: Suprya JayadevDate: December 13, 19941) Transfect cells.2) Culture cells 1-3 passages in a T-75 flask containing selection material (e.g.
Fibroblast－Cell－Systems4
3. If your result falls into any quadrant other than the "High Yield-High Viability" quadrant, refer to Appendix D, Improving Cell Yield and Viability
使用CO2恒溫搖床解決人胚腎－293－(HEK293)－細胞結團問題
人胚腎 293 (HEK293)? 細胞在重組蛋白表達中是最常見的宿主細胞。 這類細胞能夠表達大量的膜蛋白，如 G 蛋白偶聯受體? (GPCR) ，是無法在最常見的生物制藥生產宿主，如：中國倉鼠卵巢 (CHO) 細胞中作表達。 HEK293 雖然是蛋白表達的極好宿主，然而 HEK293 細胞
Transfer－of－Eukaryote－Suspension－Cultures
MaterialsFibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Disp
Handling－Fresh－Hepatocytes－in－Suspension
實驗概要Important—Please read!GIBCO? ?Fresh Hepatocytes are shipped in cold preservation medium designed to ?keep cells viable at 4°C. This shipping mediu
Dissociation－of－Cells－from－Primary－Tissue
實驗概要A ?common method to obtain single cell suspensions from primary tissue is ?enzymatic disaggregation. Expose the cells to enzymes for a minimal ?am
Immunofluorescence－/－Confocal－Microscopy－Protocol
實驗概要Immunofluorescence ?is a technique used for light microscopy with a fluorescence microscope ?and is used primarily on biological samples. This tec