Stem Cell Isolation Protocols

The starting material for de novo isolation of stem cell lines can be either normal 3.5-days post coitum (dpc) expanded blastocysts or "delayed" blastocysts. Delayed blastocysts are usually collected 4-6 days after ovariectomy. For both groups of blastocysts, tissue culture procedures are similar. The only difference is the timing of the first disaggregation, because delayed blastocysts will initially grow more slowly. - [Read De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts Protocol]

This protocol describes a method for freezing ES cells in 96-well plates after ES colonies have been isolated by picking. - [Read Freezing Embryonic Stem (ES) Cells in 96-Well Plates Protocol]

This protocol describes a method for isolating individual ES cell colonies by picking after they have grown on selective medium. - [Read Isolating Individual Embryonic Stem (ES) Cell Colonies by Picking Protocol]

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolation of early neural-fold-stage embryos. - [Read Isolating Postimplantation Embryos: Early Neural-Fold-Stage Protocol]

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause post-implantation defects. This protocol describes a method for isolating early primitive-streak-stage embryos. - [Read Isolating Postimplantation Embryos: Early Primitive-Streak-Stage Protocol]

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating early somite-stage embryos (~8.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Early Somite-Stage Protocol]

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating late primitive-streak-stage embryos (~7.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Late Primitive-Streak-Stage Protocol]

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations that cause post-implantation defects. This protocol describes a method for isolating prestreak-stage embryos (~5.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Prestreak-Stage Protocol]

This protocol describes the isolation of fibroblasts from chicken embryos. - [Read Isolation of Fibroblasts from Chicken Embryos Protocol]

Protocol for the production of completely ES cell-derived fetuses by aggregation with tetraploid embryos. Includes: Recovery of 2-cell stage embryos; Production of tetraploid embryos; Preparation of aggregation plate; Removal of Zona Pellucida; ES cells/ tetraploid embryo "SANDWICH" aggregation; Transfer of embryos. - [Read Production of Completely ES Cell-Derived Fetuses by Aggregation with Tetraploid Embryos]

Germ layers are multipotent tissues that have the ability to differentiate into various tissue types. Isolation and manipulation of germ layers is important for examining differentiation. This protocol describes a method for separating postimplantation germ layers. - [Read Separating Postimplantation Germ Layers Protocol]