LIF1005 Sigma-AldrichLeukemia Inhibitory Factor human

Human embryonic stem cells (hESC), with their ability to differentiate into all cell types in the human body, are likely to play a very important therapeutic role in a variety of neurodegenerative and life-threatening disorders in the near future. Although more than 120 different human embryonic stem cell lines have been reported worldwide, only a handful are currently available for researchers, which limits the number of studies that can be performed. This study reports the isolation, establishment and characterization of new human embryonic stem cell lines, as well as their differentiation potential into variety of somatic cell types. Blastocyst-stage embryos donated for research after assisted reproductive techniques were used for embryonic stem cell isolation. A total of 31 blastocysts were processed either for immunosurgery or direct culture methods for inner cell mass isolation. A total of nine primary stem cell colonies were isolated and of these, seven cell lines were further expanded and passaged. Established lines were characterized by their cellular and colony morphology, karyotypes and immunocytochemical properties. They were also successfully cryopreserved/thawed and showed similar growth and cellular properties upon thawing. When induced to differentiate in vitro, these cells formed a variety of somatic cell lineages including cells of endoderm, ectoderm and mesoderm origin. There is now an exponentially growing interest in stem cell biology as well as its therapeutic applications for life-threatening human diseases. However, limited availability of stem cell lines as well as financial or ethical limitations restrict the number of research projects. The establishment of new hESC lines may create additional potential sources for further worldwide and nationwide research on stem cells.

Using in situ hybridization histochemistry, we characterized the spatiotemporal gene expression patterns of leukemia inhibitory factor (LIF) and glial cell line-derived neurotrophic factor (GDNF), and their receptor components (LIFR, GFR-alpha1, RET) induced in muscle cells, intramuscular nerves, and motoneurons in the regeneration processes of both muscle cells and nerves following muscle contusion. Muscle contusion induced upregulation of GDNF and GFR-alpha1 mRNAs in Schwann cell-like cells in the intramuscular nerves and of LIFR mRNA in damaged muscle cells. LIFR, GFR-alpha1, and RET mRNA expressions in motoneurons were upregulated following muscle contusion. Muscle contusion also induced more rapid, prominent transactivations of GFR-alpha1 and RET genes in motoneurons than did sciatic nerve axotomy. These findings suggest that rapid and prominent upregulation of the receptor components for LIF and GDNF in motoneurons is important for the regeneration of intramuscular motor nerves damaged by muscle contusion.

The isolation and expansion of human neural progenitor cells have important potential clinical applications, because these cells may be used as graft material in cell therapies to regenerate tissue and/or function in patients with central nervous system (CNS) disorders. This paper describes a continuously dividing multipotent population of progenitor cells in the human embryonic forebrain that can be propagated in vitro. These cells can be maintained and expanded using a serum-free defined medium containing basic fibroblast growth factor (bFGF), leukemia inhibitory factor (LIF), and epidermal growth factor (EGF). Using these three factors, the cell cultures expand and remain multipotent for at least 1 year in vitro. This period of expansion results in a 10(7)-fold increase of this heterogeneous population of cells. Upon differentiation, they form neurons, astrocytes, and oligodendrocytes, the three main phenotypes in the CNS. Moreover, GABA-immunoreactive and tyrosine hydroxylase-immunoreactive neurons can be identified. These results demonstrate the feasibility of long-term in vitro expansion of human neural progenitor cells. The advantages of such a population of neural precursors for allogeneic transplantation include the ability to provide an expandable, well-characterized, defined cell source which can form specific neuronal or glial subtypes.