Morula-derived human embryonic stem cells

Abstract

Human embryonic stem (ES) cells are known to derive from the inner cell mass of blastocyst. Although the embryos of other developmental stages have also been used as a source for ES cells in animal models, the feasibility of obtaining ES cell lines from human morula is not known, despite being an obvious source available through assisted reproduction and preimplantation genetic diagnosis programmes. This study describes an original technique for derivation of ES cells from human morula, which enabled the establishment of eight morula-derived ES cell lines. These ES cell lines were shown to have no morphological differences from the ES cells derived from blastocysts, and expressed the same ES cell specific markers, including Oct-4, tumour-resistance antigens TRA-2-39, stage-specific embryonic antigens SSEA-3 and SSEA-4, and high molecular weight glycoproteins TRA-1-60 and TRA-1-81, detected in the same colony of morula-derived ES cells showing specific alkaline phosphatase expression. No differences were observed in these marker expressions in the morula-derived ES cells cultured in the feeder layer free medium. Similar to ES cell originating from blastocyst, the morula-derived ES cells were shown to spontaneously differentiate in vitro into a variety of cell types, including the neuron-like and contracting primitive cardiocyte-like cells.

Derivation of germ-line-competent embryonic stem cell lines from preblastocyst mouse embryos

“…These findings show that genuine pluripotent cell lines can de derived from preblastocyst stage mouse embryos, and they provide a simple and efficient alternative to the blastocyst as sources of such cells. Putative ES cells have also been obtained recently from human morulae by initially culturing the morulae underneath a feeder layer (26).…”

Defined culture conditions of human embryonic stem cells

“…April͞BAFF ͉ fibroblast growth factor ͉ serum free culture ͉ wnt H uman embryonic stem cells (hESCs) are pluripotent cells that have the potential to differentiate into the three germ layers and possibly all tissues of the human body (1)(2)(3)(4)(5)(6). hESCs were originally isolated from the inner cell mass of human embryos and can be passaged through Ͼ100 divisions in vitro (7,8).…”

Derivation of buffalo embryonic stem-like cells from in vitro-produced blastocysts on homologous and heterologous feeder cells

“…The present study show that homologous feeder layers allow efficient derivation of BuES-like cells from hatched and expanded blastocysts than heterologous feeder layers. Successful use of homologous embryonic fibroblast feeder layers to produce ES cells has been previously done in mink [19], marsupials [20], human [21][22][23], pig [24,25] and buffalo [16,17,26]. In contrast, in an early study, Piedrahita et al [27] determined that porcine embryonic fibroblasts (PEF) did not promote attachment of the porcine ICM to the feeder layer.…”