Isolation of a primate embryonic stem cell line.

Abstract

Embryonic stem cells have the ability to remain undifferentiated and proliferate indefinitely in vitro while maintaining the potential to differentiate into derivatives of all three embryonic germ layers. Here we report the derivation of a cloned cell line (R278.5) from a rhesus monkey blastocyst that remains undifferentiated in continuous passage for >1 year, maintains a normal XY karyotype, and expresses the cell surface markers (alkaline phosphatase, stage-specific embryonic antigen 3, stage-specific embryonic antigen 4, TRA-1-60, and TRA-1-81) that are characteristic of human embryonal carcinoma cells. R278.5 cells remain undifferentiated when grown on mouse embryonic fibroblast feeder layers but differentiate or die in the absence of fibroblasts, despite the presence of recombinant human leukemia inhibitory factor. R278.5 cells allowed to differentiate in vitro secrete bioactive chorionic gonadotropin into the medium, express chorionic gonadotropin a-and ,8-subunit mRNAs, and express a-fetoprotein mRNA, indicating trophoblast and endoderm differentiation. When injected into severe combined immunodeficient mice, R278.5 cells consistently differentiate into derivatives of all three embryonic germ layers. These results define R278.5 cells as an embryonic stem cell line, to our knowledge, the first to be derived from any primate species.

Molecular cloning and functional analysis of cDNA encoding a rat leukemia inhibitory factor: towards generation of pluripotent rat embryonic stem cells

“…A possible signal sequence is underlined. An asterisk shows amino acid identities between rat and mouse Thomson et al, 1995). The cells cultured for 6 days with the supernatant which contained 1000-1500 units/ ml of LIF activity derived from cells transfected with pCXN-rLIF, clearly showed positive-staining for alkaline phosphatase activity (Figure 4a and b).…”

“…In fact, mouse ES cells maintained in mouse LIF retained the ability to form germ-line chimeric mice indicating that LIF is su cient to maintain the developmental potential of ES cell lines previously isolated and maintained on feeders (Evans and Kaufman, 1981;Capecchi, 1989). Pluripotent cell lines have been derived only from preimplantation embryos of several other animals including rat (Iannaccone et al, 1994), avian (Pain et al, 1996), mink (Sukoyan et al, 1992), hamster (Doetsman et al, 1985), pig (Wheeler, 1994), bovine (First et al, 1994), zebra ®sh (Sun et al, 1995) and primate (Thomson et al, 1995), but the developmental potentials of these cell lines remain poorly characterized and they have not yet been shown to reconstitute the germ line.…”

Clonogenic analysis reveals reserve stem cells in postnatal mammals. II. Pluripotent epiblastic‐like stem cells

“…They exhibit telomerase activity, which is consistent with their extended capability for self-renewal (Liu, 2000;Pera et al, 2000;Lin et al, 2003). When released from inhibitory control in vitro, these cells will spontaneously differentiate into and exhibit phenotypic expression markers for cells of ectodermal, mesodermal, and endodermal origin (Thomson et al, 1995(Thomson et al, , 1998Shamblott et al, 1998;Pera et al, 2000). Thus, embryonic stem cells exhibit pluripotentiality, i.e., the ability of a single cell to form multiple types of tissue from all three primary germ layer lineages.…”

“…They have been isolated from the blastocyst, inner cell mass, and gonadal ridges of rodents and primates, including humans (Evans and Kaufman, 1981;Martin, 1981;Thomson et al, 1995Thomson et al, , 1998Shamblott et al, 1998;Pera et al, 2000). After isolation and growth in vitro with inhibitory agents (i.e., leukemia inhibitory factor, ESGRO, and/or fibroblast feeder layers), these cells exhibit immunological and molecular markers for undifferentiated embryonic cells (Niwa et al, 2000;Pera et al, 2000;Pesce and Scholer, 2001;Henderson et al, 2002;Cheng et al, 2003).…”

Protein profile of basal prostate epithelial progenitor cells—stage‐specific embryonal antigen 4 expressing cells have enhanced regenerative potential in vivo

“…It was demonstrated that approximately 50% of undifferentiated murine ES cells express SSEA‐1; however, less than 10% of the more differentiated cells express SSEA‐1 22. ES cells of monkeys and humans in contrast express SSEA‐3 and SSEA‐4 as markers, but not SSEA‐1 16, 23. Similar to SSEA‐1 in the mouse, human SSEA‐3 and SSEA‐4 expression correlated with the level of differentiation of ES cell lines 24.…”