ESCellCultureandManipulation
MediaHigh glucose DMEM (-pyruvate, -glutamine)20% Heat-inactivated Fetal calf serum (can vary by cell type, be sure!!)1X l-glutamine1X Penicillin/streptomycin1X Non-essential amino acids1X ribonucleosides1/100 volume ß-ME stock (stock is 7µl ß-ME in 10ml DMEM)1:1000 dilution of LIF (add 500 µl 720LIFD cell conditioned media to 500ml media)Thawing frozen vials of ES cellsThaw vial rapidly at 37°C, rinse outside of via......阅读全文
Human－Embryonic－Stem－(ES)－Cell－Protocols——Splitting－Human－ES－cells－on－MEFs
based on splitting onto one plateWarm collagenase IV split media to 37 °C in a water bath.Aspirate media off of cell culture plate.Add the following a
RNA－Isolation－From－Animal－tissue－or－cell－culture
实验概要This method is designed for most animal tissues and culture cells. For RNA isolation from fibrous tissue, follow the specialized protocol on pag
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
293细胞培养(cell－culture)技术
1、293细胞明显适应酸性环境,pH值在6.9～7.1时,可顺利贴壁生长, 换液时动作要轻。一般用高糖的DMEM培养基。2、传代：倒去废液，PBS洗一次（轻），用0.02%EDTA与0.25%Trypsin消化，生长良好细胞，培养瓶中轻摇,使之流遍所有细胞表面,即将其吸除或弃去消化30s，然后吸去，
Human－Embryonic－Stem－(ES)－Cell－Protocols——Embryonic－Bodies
Let human ES cells grow until the colonies are large and the cells are pretty piled up - about the time when you would normally split or even a day pa
Human－Embryonic－Stem－(ES)－Cell－Protocols——Media－and－Reagents
Serum Free Media for human ES cells on MEFs: can last for 7-10 daysFinal ConcentrationAmount for 250ml Stock solution80% DMEM-F12200ml20% KO Serum Rep
植物细胞培养（plant－cell－culture）技术概述
植物细胞培养技术就是为了某种目的而在细胞水平上对离体植物细胞或原生质体进行的一系列生物工艺学操作。它包括分离、培养、再生以及一系列相关的操作。就有用化合物的生产来说，它主要是指在无菌条件下通过悬浮培养植物细胞生产有用化合物的过程。理论与技术基础：植物细胞全能性、微生物液体深层发酵系统、遗传工程
Human－Embryonic－Stem－(ES)－Cell－Protocols——Matrigel－Aliquoting－and－Plating
Aliquoting Matrigel: Day one: Put the sterilized tip box (either 200 ml or 1000 ml tips), sterilized eppendorf tube container, and appropriate pipe
Differentiate－ES－cells－into－glial－cells－and－neurons
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells. ___________________ Day 1: Tryp
Growing－feederindependent－embryonic－stem－cells§
We use feeder-independent ES cell lines derived from the 129/Ola strain of mice (Nichols et al., Development 110, p.1341, 1990). These cells are easy
原代神经细胞培养方法－Neuron－Cell－Culture
1. Preparation of coverslips1.1- Mass cultureOur standard mass cultures are plated on astrocytes. Those, in turn, are plated on glass coverslips pre-
核移植胚胎干细胞的印迹基因甲基化研究
核移植来源的胚胎干细胞(NTES cells)在以干细胞为基础的细胞治疗中扮演着非常重要的角色，得到全能性良好且表观遗传修饰正常的核移植胚胎干细胞是解决治疗性克隆安全问题的重要前提。DNA甲基化修饰在基因表达和印迹基因的表达中起非常重要的作用，两步法克隆可能存在的不完全重编程问题很可能存在于印
Bacteria－Growth－and－Culture－Bacteria－Growth－and－Culture
Flagella and motilitymonotrichous flagella - the bacterial cell has a single flagellaperitrichous flagella - the bacterial cell has several flagella w
Differentiate－ES－cells－into－cardiac－myocytes
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells. ____________________ Day 1: Try
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
Lactobacillus－culture
OverviewGeneral overview and guidelines on how to grow up a culture of LactobacillusMaterialsMRS broth (difco)MRS agar (difco)anaerobic conditionsProc
Construction－and－Manipulation－of－LargeInsert－Bacterial－Clone－Libraries
Acknowledgements The organizer of the workshop acknowledges Dr. Murray Milford, Professor and Interim Head, and Dr. Mark Hussey, Professor and Interim
Isolation,－Culture,－and－Differentiation－of－Progenitor－Cells2
Slide Coating13. Just before use, dilute the poly-D-lysine stock solution 1:50 in H2O. Add 0.2 mL of diluted poly-D-lysine to each well of an eight-we
Differentiate－ES－cells－into－cystic－embryoid－bodies
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.______________Day 1: Trypsinized th
Culture－of－Endometrial－Explants
Culture of Endometrial Explants and Peri-implantation Conceptuses to Monitor Synthesis and Secretion of Proteins and Prostaglandins P. J. Hansen
Tissue－Culture－Media
We use two different kinds of media. Most cells are grown in DMEM. A few lymphoid cell lines are grown in RPMI. Cells grown in DMEM must be grown in
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
Acetobacter－Xylinum－Culture
OverviewGeneral guidelines on how to grow up a culture of Acetobacter xylinum, ATCC strain 53582 .Preparation of Acetobacter MediaTo prepare ∼500 ml o
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). Ho
Bacteria－Culture－Protocol
Bacteria Culture ProtocolBy 徐晓政1、TBS Medium Preparation:Prepare 1L of TBS medium contains:Tryptone 12gYeast extract 24gNaCl 5gSodium Succinate 5gGlyce
Electroporation－of－ES－cells
Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for
KARYOTYPING－ES－CELLS
An actively growing culture of cells is required, i e 2 - 3 d ES cell culture. The total number of cells needs to be between 106 - 107 cells. N B Re
细胞培养（cell－culture）的一些常见问题与解答2
6、血清的有关问题：新生牛血清：新出生牛5天内取血制成小牛血清：小牛出生16周以内采血制成。胎牛血清：（进口血清）要求剖腹取胎制成。国内厂家往往不能做到，经常把出生2天以内采血称为胎牛血清。根据血清的生产工艺及血红蛋白、内毒素含量又分为：（1）特级胎牛血清：40纳米过滤，内毒素含量≤10EU，血红蛋
细胞培养（cell－culture）的一些常见问题与解答2
6、血清的有关问题：新生牛血清：新出生牛5天内取血制成小牛血清：小牛出生16周以内采血制成。胎牛血清：（进口血清）要求剖腹取胎制成。国内厂家往往不能做到，经常把出生2天以内采血称为胎牛血清。根据血清的生产工艺及血红蛋白、内毒素含量又分为：（1）特级胎牛血清：40纳米过滤，内毒素含量≤10EU，血红蛋
细胞培养（cell－culture）的一些常见问题与解答1
1、细胞传代消化时所用胰酶浓度越高越好吗？不见得！因为胰蛋白酶溶液的消化能力是和溶液的pH、温度、胰酶浓度及溶液中是否含有Ca++， Mg++离子和血清等因素有关。通常情况下，pH 8.0， 温度37 ℃其作用能力最强。另外，钙、镁离子及血清均能大大降低其消化能力。我们每次进行传代消化前，一