-
[show abstract]
[hide abstract]
ABSTRACT: Recent progress in somatic cell nuclear transfer (SCNT) provides the evidence for the presence of reprogramming factors in
human embryonic stem cells (hESC). Hybrid hESC with donor human somatic nuclei have been established, but the resulting hybrid
hESC contained DNA originating from both hESC and donor somatic cells. There is still no method to completely replace the
hESC nuclei by the nuclei of somatic cells and to obtain the pure colonies of hESC with donor genotype. We present here the
original technology, which is based on enucleation of h ESC and their fusion with the adult somatic cells, resulting in the
establishment of individual-specific hESC with the genotype of the donor somatic cells. The resulting constructs was demonstrated
to have the “stemness” of hESC and the genotype of the donor somatic cells. So this “Stembrid technology,” may be used for
the construction of patient-specific hESC.
Stem Cell Reviews and Reports 04/2012; 2(4):297-299. · 3.74 Impact Factor
-
Katherine Amps,
Peter W Andrews,
George Anyfantis,
Lyle Armstrong,
Stuart Avery,
Hossein Baharvand,
Julie Baker,
Duncan Baker,
Maria B Munoz,
Stephen Beil, [......],
Kazutoshi Takahashi,
Timo Tuuri,
Parvathy Venu,
Yuri Verlinsky,
Dorien Ward-van Oostwaard,
Daniel J Weisenberger,
Yue Wu,
Shinya Yamanaka,
Lorraine Young,
Qi Zhou
[show abstract]
[hide abstract]
ABSTRACT: The International Stem Cell Initiative analyzed 125 human embryonic stem (ES) cell lines and 11 induced pluripotent stem (iPS) cell lines, from 38 laboratories worldwide, for genetic changes occurring during culture. Most lines were analyzed at an early and late passage. Single-nucleotide polymorphism (SNP) analysis revealed that they included representatives of most major ethnic groups. Most lines remained karyotypically normal, but there was a progressive tendency to acquire changes on prolonged culture, commonly affecting chromosomes 1, 12, 17 and 20. DNA methylation patterns changed haphazardly with no link to time in culture. Structural variants, determined from the SNP arrays, also appeared sporadically. No common variants related to culture were observed on chromosomes 1, 12 and 17, but a minimal amplicon in chromosome 20q11.21, including three genes expressed in human ES cells, ID1, BCL2L1 and HM13, occurred in >20% of the lines. Of these genes, BCL2L1 is a strong candidate for driving culture adaptation of ES cells.
Nature Biotechnology 11/2011; 29(12):1132-44. · 29.50 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: There are several sources from which human embryonic stem cell (hESC) lines can be generated: surplus embryos after in vitro fertilization procedures, one- and three-pronuclear zygotes, early arrested or highly fragmented embryos that have reached the blastocyst stage, or otherwise chromosomally or genetically abnormal embryos after preimplantation genetic diagnosis (PGD). We report on the efficiency of establishing hESC lines from blastocysts with proven meiotic or mitotic errors after sequential testing of both polar bodies and blastomere analysis on day 3. The success rate of establishing hESC lines originating from blastocysts carrying a meiotic error was as low as 2.4% and differed significantly from the success rate of establishing hESC lines originating from blastocysts with balanced meiotic errors (21.6%) or mitotic errors (after sequential testing (9.1%) and after blastomere testing alone (12.2%)). This suggests that it may be reasonable to apply sequential PGD prior to the initiation of hESC culture. Information about the karyotype may in the future help refine the methods and possibly improve the efficiency by which hESC lines are derived from embryos with prezygotic abnormalities. Additionally, it may in general prove very difficult to obtain abnormal hESC lines for scientific study from aneuploid PGD embryos, which will limit our ability to study the biological consequences of chromosomal abnormalities. Furthermore, the success rates for generating aneuploid cell lines originating from fertilized oocytes carrying a prezygotic nondisjunction error seem to mirror the miscarriage rates during pregnancy of embryos carrying such errors.
Stem cell research 02/2009; 2(1):78-82. · 3.39 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: At least 50-60% of oocytes derived from IVF procedures are chromosomally abnormal due to meiotic I or II errors. Through the use of polar body and blastomere diagnosis, euploid embryos suitable for transfer can be identified. Those embryos that are aneuploid are usually discarded, or otherwise can be used to generate chromosomally abnormal human embryonic stem cell (hESC) lines. The authors' centre has one of the largest repositories of hESC lines with genetic and chromosomal disorders generated from preimplantation genetic diagnosis (PGD) abnormal embryos. The results, studying hESC lines derived from PGD abnormal zygotes, imply that aneuploidies resulting from meiotic non-disjunction have a greater impact on viability of cells of the human embryos than those originating from post-zygotic mitotic non-disjunction.
Reproductive biomedicine online 02/2009; 18(1):120-6. · 2.04 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Recent progress in somatic cell nuclear transfer (SCNT) provides the evidence for the presence of reprogramming factors in human embryonic stem cells (hESC). Hybrid hESC with donor human somatic nuclei have been established, but the resulting hybrid hESC contained DNA originating from both hESC and donor somatic cells. There is still no method to completely replace the hESC nuclei by the nuclei of somatic cells and to obtain the pure colonies of hESC with donor genotype. We present here the original technology, which is based on enucleation of h ESC and their fusion with the adult somatic cells, resulting in the establishment of individual-specific hESC with the genotype of the donor somatic cells. The resulting constructs was demonstrated to have the "stemness" of hESC and the genotype of the donor somatic cells. So this "Stembrid technology," may be used for the construction of patient-specific hESC.
Stem Cell Reviews and Reports 02/2006; 2(4):297-9. · 3.74 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Somatic cell nuclear transfer (SCNT) provides the basis for the development of patient-specific stem cell lines. Recent progress in SCNT suggested the presence of reprogramming factors in human embryonic stem (hES) cells, although no method is currently available for replacement of nuclei of hES cells by somatic cell nuclei. An original technique has been developed, involving the fusion of different types of somatic cells with hES cells, which allowed a complete replacement of the nuclei of hES cells by nuclei of somatic cells. The resulting 'cybrids' were demonstrated to have the genotype of the donor somatic cells and 'stemness' of the recipient hES cells. However, the colonies isolated from the resulting fusion contained a mixture of these cybrid cells with the cells with the recipient nuclei, as well as hybrid cells containing both donor and recipient nuclei, so future purification will be necessary before the technique can be considered for future practical application.
Reproductive biomedicine online 02/2006; 12(1):107-11. · 2.04 Impact Factor
-
[show abstract]
[hide abstract]
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.
Reproductive biomedicine online 01/2005; 9(6):623-9. · 2.04 Impact Factor
