Establishment, Culture and Freezing of Human and Mouse Embryonic Stem Cells: a Protocol Guide

Freezing and Thawing Human Embryonic Stem Cells

Since James Thomson et al developed a technique in 1998 to isolate and grow hES in culture, freezing cells for later use and thawing and expanding cells from a frozen stock have become important procedures performed in routine hES cell culture. Since hES cells are very sensitive to the stresses of freezing and thawing, special care must taken. Here we demonstrate the proper technique for rapidl...

Improvement of Expression of α6 and β1 Integrins by the Co-culture of Adult Mouse Spermatogonial Stem Cells with SIM Mouse Embryonic Fibroblast Cells (STO) and Growth Factors

Differentiation of Mouse Embryonic Stem Cells into Hematopoietic Cells

Purpose: Differentiation of Mouse embryonicstem cells into Hematopoietic cells. Materials and Methods: In this study, we used EB formation system for Hematopoietic differentiation of mouse embryonic stem cell (Royan B1) in suspension culture. EBs cultured in medium with Hematopoietic inducer cytokines (SCF, TPO, GMCSF, IL3, Flt3 and EPO) .presence of hematopoietic differentiated cell assessed ...

Extract of mouse embryonic stem cells induces the expression of pluripotency genes in human adipose tissue-derived stem cells

Objective(s): In some previous studies, the extract of embryonic carcinoma cells (ECCs) and embryonic stem cells (ESCs) have been used to reprogram somatic cells to more dedifferentiated state. The aim of this study was to investigate the effect of mouse ESCs extract on the expression of some pluripotency markers in human adipose tissue-derived stem cells (ADSCs). Materials and Methods: Human A...

Co-culture of Mouse Embryonic Stem Cells with Sertoli Cells Promote in vitro Generation of Germ Cells

  Objective(s): Sertoli cells support in vivo germ cell production; but, its exact mechanism has not been well understood. The present study was designed to analyze the effect of Sertoli cells in differentiation of mouse embryonic stem cells (mESCs) to germ cells.   Materials and Methods: A fusion construct composed of a Stra8 gene promoter and the coding region of enhanced green fluorescence p...

Culture of human embryonic stem cells on human and mouse feeder cells.

This chapter describes the methods we use to maintain and expand undifferentiated human embryonic stem (hES) cells on human and mouse feeder cells. All of the available hES cells have been derived and propagated on primary mouse embryonic fibroblasts as feeder cells that have been mitotically inactivated. We found that hES cells can be successfully cultured on selected human feeder cells, such ...