细胞技术

DescriptionAllows one to test the capability of adherent cells to survive and replicate following insult with chemicals or radiation. Procedure1. Grow the cells in regular media to 70% confluence and ...

Materials Monolayer cultures grown in 75 mm culture flasks (Cells from Exercise 11.4 may be used or cultures of tetrahymena yeast or algae may be used.)H-thymidine with at least 4 µc/ml 0.36 c/mMPhosp ...

DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells noble agar assay can be utilized. Noble agar is used to ...

-Use sterile technique and sterile solutions throughout this method.- 1. Grow a starter culture at 30 C with shaking (250 rpm) until it reaches saturation. ***It is important to make sure all strains ...

Preparation of cell lysate: 1. For cells in suspension grow in DMEM with 10% FCS and then the day before you do the experiment spin the cells down (1 K for 5 min) and obtain a concentration of cells a ...

Introduction Mature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR). The most immediate consequence of TCR activation is the initiation of signa ...

OverviewThis assay uses a simple apparatus to study chemotaxis of leukocytes or other migratory cells. The apparatus consists of two multi-well chambers separated by a filter containing pores of unif ...

ADSC isolation culture and cloningRat amnion membrane is mechanically separated from the chorion of embryonic day 18.5 Sprague�Dawley rat embryos The tissue is washed extensively with phosphate-buffer ...

Introduction A modification of the basic immunofluorescent staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surface molecules and intracellular antigens at t ...

The following protocol is for detection of human placental alkaline phosphatase (AP) in cultured cells. Human placental AP is heat stable unlike other AP proteins typically expressed by cells. A few c ...

Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.____________________Day 1: Trypsinized the cells as for normal passaging until the co ...

AP Buffer: 100 mM Tris-HCl pH 9.5 100 mM NaCl 10 mM MgCl2 PTM: PBS 0.1% Tween-20 2 mM MgCl2 Since this is generally done in conjunction with lacZ staining embryo/tissue is usually fixed as per lacZ st ...

Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.___________________Day 1: Trypsinized the cells as for normal passaging until the col ...

Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.______________Day 1: Trypsinized the cells as for normal passaging until the colonies ...

Media UsedTo prepare 100 ml mediumDMEM 80 ml FCS 15 ml Non-essential amino acids (100x) 1 ml Pen/strep (5000 1U/ml 5000 ug/ml) 1 ml L-Glutamine 200 mM 1 ml Nucleosides stock (100x) 1 ml BME 0.1M 0.2 m ...

Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for 1-2 electroporations. Procedure1. Change medium on ...

Needed:ES cell freezing medium (2x)2x ES cell freezing medium should be made up fresh each time it is to be used and should comprise freshly prepared 60% DMEM+ 20% FCS and 20% DMSO (Sigma Cat. No.D-58 ...

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 to maintain and significantly reduce the amount of ti ...

IntroductionHuman embryonic stem (hES) cells are pluripotent stem cells derived from pre-implantation embryos that can be maintained and expanded in an undifferentiated state or induced to differentia ...

MEF Conditioned mediumPlate MEFs (p4 or p5) onto gelatin-coated plates at a density of 1x106 cells/10cm culture dish. The next day wash cells with PBS and add 10ml normal hESC medium containing 4ng/ml ...