Stem cell lines

The blastocyst is an early embryonic stage in mammalian development. Murine blastocysts can be harvested at day 3.5 p.c. Their inner cell mass contains embryonic stem cells. Multiple murine embryonic stem cell lines have been established. Embryonic stem cells carrying genetically engineered mutations are injected into blastocysts, which are subsequently implanted into pseudopregnant foster mothers. 

Buzanska, L., Stachowiak, E., Stachowiak, M. and Domanska-Janik, K. Neural stem cell line derived from human umbilical cord blood - morphological and functional properties. J. Neurochem. 85(Suppl. 2) 33, 2003. 

The murine hematopoietic stem cell line Myl-D7 spontaneously differentiate along the lymphoid, myeloid and erythroid lineages. Myl-7 cells shows a strict stromal dependence for growth of self-renewing stem cells and express high levels of CSF-1 receptor (Itoh et al., 1996). We used this cell line to analyze the function of CSF-1 in maintaining multipotent cells. In an other attempt to characterize unknown factors that could sustain stem cells we... 

We next analyzed the effects of the CSF-1 isoforms on the development of Myl-D7, a strictly stroma-dependent stem cell line (Itoh et al, 1996). At least 51% of Myl-D7 cells express among other multilineage markers high levels of CSF-1 receptors (data not shown). 

Figure 6. Proliferation of the murine Myl-D7 stem cell line is stimulated efficiendy only by membrane-bound CSF-1. 2-10 Myl-D7 cells were cultured on MMCE cells expressing the different CSF-1 isoforms in 48-well plates. Cloning efficiencies were determined weekly and Myl-D7 clones (>10 cells) were transferred onto new MMCE cells. Values of Myl-D7/MS5 cocultures were set to 100% at each time point (data not shown). Results are expresses as percentage of control Myl-D7/MS5 cultures and are mean values +SD (four experiments). After 3 weeks in coculture a) no difference was found in the proliferation rates of cells stimulated by either wildtype- or soluble CSF-1 (P>.05) b) mean values for populations stimulated by membrane-bound- or soluble CSF-1 were significantly different (P<.001).

The stem cell line Myl-D7 not only expresses differentiation markers of all lineages but is dependent for growth on self-renewing stem cells within the Myl-D7 clone which spontaneously differentiates along the lymphoid, myeloid, or erythroid direction (Itoh et ah, 1996). Supernatant of MS5 induces short term growth of Myl-D7 cells. This may indicate that at least soluble stem cell growth factors are secreted by MSS cells. 

We used the stroma dependent Myl-D7 cell line to define unknown stem cell factors secreted by MSS stromal cells. CSF-1 was identified as a major growth factor component of the MSS supernatant by biochemical analysis. Recombinant CSF-1 induced short-term growth (3 days) of Myl-D7 to a similar extent as MSS CM. Furthermore, growth of the parental Myl-D7 clone in MSS CM could be inhibited by addition of an a-CSF-1 antibody, indicating that at least one of the active molecules is CSF-1. Thus we conclude that CSF-1 is a potent stimulator of the limited proliferation of the Myl-D7 stem cell line (Heberlein et al., 2006). 

Heberlein C, Friel J, Itoh K, Medlock E, Li L, Nakayama N, Stocking C, Geldmacher M, Ostertag W, 2006. Involvement of CSF-1 in generating a stroma-independent hematopoietic stem cell line. J Cell Physiol 206 556... 

Harrison D. E. (1979). Mouse erythropoietic stem cell lines function normally 100 months loss related to number of transplantations, Mech. Ageing Dev., 9, 427-433. 

Embryonic stem cell lines derived from hrnnan blastocysts. Science 1998 282 1145 7. Thomson, J.A. et al. Isolation of a primate embryonic stem cell line. Proc.Nad. Acad. Sci. USA 92, 7844-7844 (1995). 

Pollard SM, Yoshikawa K, Clarke ID et al (2009) Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. Cell Stem Cell 4 568-580... 

Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM. Embryonic stem cell lines derived from human blastocysts. Science 1998 282 1145-1147. 

Cowan CA, Klimanskaya I, McMahon J, Atienza J, Witmyer J, Zucker IP, Wang S, Morton CC, McMahon AP, Powers D, Melton DA. Derivation of embryonic stem-cell lines from human blastocysts. N Engl J Med 2004 350 1353-1356. 

The EST has continued to be a popular test subject to further optimization, especially through the addition of alternative pathways of differentiation and the addition of molecular markers of effects. Besides cardiac muscle differentiation, other cell types such as neurons (29-31), osteoblasts (32), adipocytes (33), and hepato-cytes (34) have been generated in dedicated differentiation protocols, providing additional opportunities for testing the interference of differentiation pathways with chemical exposures. The addition of transcriptomics approaches to measure differential gene expression, both as influenced by the differentiation process as well as due to chemical exposure, has proven informative and may enhance the predictability of EST assays (35). The extension of this concept to human embryonic stem cell lines is still in its infancy, but has opened a new realm of options that bypasses the need for interspecies extrapolation (36). 

Complete medium for culture of the undifferentiated ES-D3 embryonic stem cell line Dulbecco s Modified Eagle s Medium (DMEM), 20% Fetal bovine serum (FBS), 1% Nonessential Amino Acids (NEAA) (Gibco, Gaithersburg, MD), 2 mM L-Glutamine, 1% 5,000 lU/ml Penicillin/5,000 pg/ml Streptomycin, 0.1 mM P-Mercaptoethanol. 

Wakayama, T., V. Tabar, I. Rodriguez, A.C. Perry, L. Studer, and P. Mombaerts, Differentiation of embryonic stem cell lines generated from adult somatic cells by nuclear transfer. Science, 2001. 292(5517) 740-3. 

Brown, A.B., Yang, W., Schmidt, N.O., Carroll, R., Leishear, K.K., Rainov, N.G., Black, P.M., Breakefield, X.O., Aboody, K.S. (2003). Intravascular delivery of neural stem cell lines to target intracranial and extracranial tumors of neural and non-ncural origin. Hum Gene Ther, 14, 1777-85. 

Thomson, J.A., Itskovitz-Eldor, J., Shapiro, S.S., Waknitz, M.A., Swiergiel, J.J., Marshall, V.S., Jones, J.M. (1998). Embryonic stem cell lines derived from human blastocysts. Science. 282, 1145-7. Erratum in (1998) Science, 282, 1827. 

Reubinoff, B.E., Pera, M.F., Fong, C.Y., Trounson, A., Bongso, A. (2000). Embryonic stem cell lines from human blastocysts somatic differentiation in vitro. Nat Biotechnol, 18, 399-404. Erratum in (2000) Nat Biotechnol. 18, 559. 

Wakayama, T. (2006). Establishment of nuclear transfer embryonic stem cell lines from adult somatic cells by nuclear transfer and its application. Ernst Sobering Res Found Workshop, 60, 111-23. 

Grompe, M., Overturf, K., Al-Dhalimy, M., and Finegold. M. Serial transplantation reveals stem cell line regenerative potential in parechymal mouse hepatoctes. Hepatology 24, 256A 1996. 

While the hydra is almost immortal as a result of the continuous differentiation of its stem cell lines, other small invertebrates follow a very different course of development. Both the rotifers and the annelid worms (Fig. 1-14) tend to have a constant number of cells in the adult body. The entire developmental program is specified genetically in strict detail. 

Doetschman, T. C., Eistetter, H., Katz, M., Schmidt, W. and Kemler, R. (1985) The in vitro development of blastocyst-derived embryonic stem cell lines formation of visceral yolk sac, blood islands and myocardium. JEmbryol Exp Morphol 87, 27—45. 

Abbondanzo, S.J., Gadi, I., and Stewart, C.L. (1993) Derivation of embryonic stem cell lines. Methods Enzymol. 225, 803-823. 

Hwang WS, Ryu YJ, Park JH, Park ES, Lee E, Koo, JM, Jeon HY, Lee BC, Kang SK, Kim SJ, Ahn C, Hwang JH, Park KY, Cibelli JB, Moon SY (2004), Evidence of a pluripotent human embryonic stem cell line derived from a cloned blastocyst, Science 303 1669-1674. 

Embryonic stem cell The embryonic stem cell line is typically derived from a male agouti 129/terSv embryo. The cell line must be maintained with meticulous culture procedures to retain normal karyotype and an undifferentiated state. 

Irradiated embryonic fibroblast feeder cell layer to be used for growing the embryonic stem cell line G418 resistant embryonic fibroblast cells are derived from embryos that carry a targeted disruption such as the 3-2 microglobulin gene (9). 

First human embryonic stem cell line derived (hESC)... 

Klimanskaya I, Chung Y, Becker S, Lu S-J, Lanza R. Human embryonic stem cell lines derived from single blastomeres. Nature 2006 444 481 1. 

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