Cloning Stem Cells最新文献

{"title":"Cells derived from human umbilical cord blood support the long-term growth of undifferentiated human embryonic stem cells.","authors":"Xiangcan Zhan,&nbsp;Christine Hill,&nbsp;Cory F Brayton,&nbsp;Michael J Shamblott","doi":"10.1089/clo.2007.0087","DOIUrl":"https://doi.org/10.1089/clo.2007.0087","url":null,"abstract":"<p><p>Various types of human cells have been tested as feeder cells for the undifferentiated growth of human embryonic stem cells (hESCs) in vitro. We report here the successful culture of two hESC lines (H1 and H9) on human umbilical cord blood (UCB)-derived fibroblast-like cells. These cells permit the long-term continuous growth of undifferentiated and pluripotent hESCs. The cultured hESCs had normal karyotypes, expressed OCT-4, SSEA-4, TRA-1-60, and TRA-1-81, formed cystic embryonic body in vitro and teratomas in vivo after injected into immunodeficient mice. The wide availability of clinical-grade human UCB makes it a promising source of support cells for the growth of hESC for use in cell therapies.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"513-22"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2007.0087","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27702282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The developmental potential of parthenogenetic and somatic cell nuclear-transferred rat oocytes in vitro.","authors":"Shigetoshi Mizumoto,&nbsp;Yoko Kato,&nbsp;Yukio Tsunoda","doi":"10.1089/clo.2008.0017","DOIUrl":"https://doi.org/10.1089/clo.2008.0017","url":null,"abstract":"<p><p>We examined the optimal conditions for somatic cell nuclear transfer (SCNT) in rat oocytes. First, we compared the effects of two types of inhibitors of spontaneous activation, MG132 and demecolcine, on the developmental potential of parthenogenetic oocytes. The potential of activated oocytes to develop into blastocysts significantly decreased 2 h after oocyte recovery (77% vs. 7%). The developmental potential of oocytes preserved in MG132-supplemented medium for 1 to 4 h was high (62% to 77%), but the potential of those preserved in demecolcine-supplemented medium for 3 and 4 h was low (77% vs. 41% and 37%, respectively). Second, the effect of the duration of parthenogenetic activation on the developmental potential was examined. When oocytes preserved in MG132 for 4 h were treated with 10 mM strontium for 5 or 6 h, the potential of activated oocytes to develop into blastocysts was high (78% and 70%, respectively). Using the optimal conditions for parthenogenetic activation, we examined the potential of rat enucleated oocytes receiving cumulus cells to develop into blastocysts. In contrast to parthenogenotes, the potential of SCNT rat oocytes to develop into blastocysts was low (2%) even if then oocytes were treated with the histone deacetylation inhibitor trichostatin A. The reason for the low developmental potential of rat SCNT oocytes is discussed.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"453-9"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0017","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27872920","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Germinative testicular cells and bone marrow mononuclear cells transplanted to a rat model of testicular degeneration.","authors":"Marilise M Horn,&nbsp;Ana H Paz,&nbsp;Marcos E Duarte,&nbsp;Guiherme Baldo,&nbsp;Maria C Belardinelli,&nbsp;Ursula Matte,&nbsp;Elizabeth O Cirne Lima,&nbsp;Eduardo P Passos","doi":"10.1089/clo.2008.0004","DOIUrl":"https://doi.org/10.1089/clo.2008.0004","url":null,"abstract":"<p><p>The aim of this study was to evaluate the ability of rat mononuclear bone marrow cells to recover testis cell associations and multiplication in busulfan-treated rats, and to compare these data to germinative testicular cell transplant. The germinative testicular cells were obtained by the trypsin digestion method, and bone marrow cells were harvested from femurs and tibias, and purified using by Ficoll gradient. Cell transplantation was performed by the injection of cells through the efferent ducts into the rete testis in busulfan-treated animals. Fifteen days after transplantation, the recipient rats were sacrificed and the testes were collected and analyzed by histology (hematoxilin-eosin and DAPI staining). Results demonstrated that germ cells transplantation promoted cellular reorganization of seminiferous epithelium 15 days later. On the other hand, no improvement in spermatogenesis regeneration was found after heterologous mononuclear bone marrow cell transplantation.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"543-6"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27678939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization, chromosomal assignment, and role of LIFR in early embryogenesis and stem cell establishment of rabbits.","authors":"Ana Paula Catunda,&nbsp;Elen Gócza,&nbsp;Bogdan V Carstea,&nbsp;Laszlo Hiripi,&nbsp;Helene Hayes,&nbsp;Claire Rogel-Gaillard,&nbsp;Maud Bertaud,&nbsp;Zsuzsanna Bosze","doi":"10.1089/clo.2008.0023","DOIUrl":"https://doi.org/10.1089/clo.2008.0023","url":null,"abstract":"<p><p>Leukemia inhibitory factor (LIF) is a multifunctional cytokine with an important role during early embryonic development, implantation, and as an inhibitor of murine embryonic stem cell differentiation. It exerts its effects by binding to the leukemia inhibitory factor receptor, a heterodimer of two transmembrane proteins, the specific leukemia inhibitory factor receptor subunit, and the common gp130. A partial cDNA clone coding for the membrane-bound form of the specific rabbit leukemia inhibitory factor receptor was isolated from the genital ridge of 13.5 days postcoitum fetus. Fluorescent in situ hybridization analysis revealed that the rabbit leukemia inhibitory factor receptor gene is located on chromosome OCU11p11.1. It has been shown that the membrane-bound rabbit leukemia inhibitory factor receptor mRNA is expressed during embryo implantation but not at earlier developmental stages. Rabbit embryonic stem cell-like line establishment is improved in the presence of LIF, and those cells express both leukemia inhibitory factor and its receptor. The withdrawal of leukemia inhibitory factor results the differentiation of embryonic stem cell-like cells to beating myocardial-like cells. Our findings suggest that the self-renewal mechanism is similar in mouse and rabbit embryonic stem cells, and expands our knowledge on the role of the LIF-LIFR signal pathway in early rabbit embryogenesis and rabbit embryonic stem cell establishment.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"523-34"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0023","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27822330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cotransfer of parthenogenetic embryos improves the pregnancy and implantation of nuclear transfer embryos in mouse.","authors":"Qinggang Meng,&nbsp;Minkang Wang,&nbsp;Claudia Ana Stanca,&nbsp;Szilard Bodo,&nbsp;Andras Dinnyes","doi":"10.1089/clo.2008.0003","DOIUrl":"https://doi.org/10.1089/clo.2008.0003","url":null,"abstract":"<p><p>The majority of somatic cell nuclear transfer (SCNT) clones dies in the peri- or postimplantation period. Improvement of the full-term healthy pregnancy rates is a key issue for the economical viability and animal welfare profile of SCNT technology. In this study the effects of cotransfer of parthenogenetic or fertilized embryos on the pregnancy and implantation of SCNT mouse embryos have been investigated. SCNT embryos were produced by transferring cumulus cell nuclei into enucleated B6D2F1 mouse oocytes, whereas parthenogenetically activated (PA) and fertilized embryos were derived from ICR mice by artificial activation with strontium and in vivo fertilization, respectively. SCNT embryos were inferior in their developmental capacity to blastocyst compared to either PA or fertilized embryos. SCNT embryos were transferred alone (SCNT), or cotransferred with two to three PA (SCNT + PA) or fertilized (SCNT + Fert) embryos into the oviducts of an ICR recipient. Both pregnancy and implantation rates originating from clones in the SCNT + PA group were significantly higher than those of SCNT group (p < 0.05). The weight of placentas of clones derived from SCNT, SCNT + PA, or SCNT + Fert was in all cases significantly higher than that of fertilized controls (p < 0.001). Most of the clones derived from SCNT embryos cotransferred with PA or fertilized embryos survived to adulthood and were fertile and healthy according to histopathological observations. Our results demonstrate in mouse that cotransfer of PA embryos improves the pregnancy and implantation of SCNT embryos without compromising the overall health of the resulting clones.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"429-34"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27641870","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reversible membrane permeabilization of mammalian cells treated with digitonin and its use for inducing nuclear reprogramming by Xenopus egg extracts.","authors":"Kei Miyamoto,&nbsp;Teruyoshi Yamashita,&nbsp;Tomoyuki Tsukiyama,&nbsp;Naoya Kitamura,&nbsp;Naojiro Minami,&nbsp;Masayasu Yamada,&nbsp;Hiroshi Imai","doi":"10.1089/clo.2008.0020","DOIUrl":"https://doi.org/10.1089/clo.2008.0020","url":null,"abstract":"<p><p>Plasma membranes can be reversibly permeabilized by Streptolysin O. The permeabilized cells can be reprogrammed and partially dedifferentiated in the cell-free system from egg extracts. However, the permeabilizing activity of Streptolysin O is not stable, and therefore it is difficult to control its activity. An alternative method for reversible permeabilization is useful for establishing a cell-free system. Here, we used a nonionic detergent, digitonin, for permeabilization. A low concentration of digitonin induced reversible permeabilization of the plasma membrane in bovine, mouse, and porcine somatic cells. The permeabilized cells were treated with Xenopus laevis egg extracts. The treated cells showed exchange of nuclear proteins from extracts such as incorporation of Xenopus-specific histone B4 and Lamin LIII into nuclei. After resealing of the membrane, the cells showed upregulation of OCT4, SOX2, and NANOG expression. Our results suggest that reversible permeabilization with digitonin can be used to induce nuclear reprogramming and to activate pluripotent genes by a cell-free system.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"535-42"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27872921","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hand-made cloned buffalo (Bubalus bubalis) embryos: comparison of different media and culture systems.","authors":"Riaz A Shah,&nbsp;Aman George,&nbsp;Manoj K Singh,&nbsp;Dharmendra Kumar,&nbsp;Manmohan S Chauhan,&nbsp;Radhaysham Manik,&nbsp;Prabhat Palta,&nbsp;Suresh K Singla","doi":"10.1089/clo.2008.0033","DOIUrl":"https://doi.org/10.1089/clo.2008.0033","url":null,"abstract":"<p><p>Hand-made cloning (HMC) has proved to be an efficient alternative to the conventional micromanipulator-based technique in some domestic animal species. This study reports the development of an effective culture system for in vitro culture of zona-free cloned buffalo (Bubalus bubalis) embryos reconstructed using adult skin fibroblast cells as nucleus donor. Cleavage and blastocyst rates observed were 52 and 0% in modified Charles Rosenkrans 2 (mCR2), 61 and 4.6% in modified Synthetic Oviductal Fluid (mSOF), and 82 and 40.3% in Research Vitro Cleave (RVCL; Cook, Australia) medium, respectively. Similarly, higher blastocyst rates (24.5 +/- 4.1%) were observed when zona-free parthenotes were cultured in RVCL medium. Culturing zona-free cloned buffalo embryos on flat surfaces (FS) yielded significantly higher (p < 0.05) blastocyst rates than Well of the Wells (WOW) or microdrops (MD). Furthermore, development in WOW was found to be significantly better than MD culture. The quality of HMC blastocysts was examined using differential staining. This study establishes the application of zona-free nuclear transfer procedures for the production of hand-made cloned buffalo embryos and the development of efficient culture system and appropriate media requirements for enhancing their preimplantation development.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"435-42"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0033","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27682807","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Treatment with proteasome inhibitor MG132 during cloning improves survival and pronuclear number of reconstructed rat embryos.","authors":"Noriaki Nakajima,&nbsp;Tomo Inomata,&nbsp;Junya Ito,&nbsp;Naomi Kashiwazaki","doi":"10.1089/clo.2008.0038","DOIUrl":"https://doi.org/10.1089/clo.2008.0038","url":null,"abstract":"<p><p>In several mammalian species including rats, successfully cloned animals have been generated using somatic cell nuclear transfer (SCNT). However, in the case of rats, additional treatment with MG132, a proteasome inhibitor, before enucleation of oocytes seems to be required for successful cloning because ovulated rat oocytes are spontaneously activated, and hence, their suppression is the key to successful cloning. A previous study on rats demonstrated that matured oocytes potentially possess lower cytostatic factor (CSF) activity compared to mouse oocytes, resulting in a low incidence of premature chromosome condensation in the reconstructed embryos after SCNT. It is known that mice having more than two pronuclei are generally observed in nuclear-transferred oocytes after induction of premature chromosome condensation, which implies successful reprogramming. This leads us to the hypothesis that MG132 treatment affects not only the inhibition of spontaneous activation but also the reprogramming and developmental ability of reconstructed rat embryos. If so, prolonged MG132 treatment during and/or after SCNT may further improve the survivability. However, the effect of MG132 treatment on reconstructed embryos after SCNT has been very limited in rats and other species. We show here that prolonged MG132 treatment during and after SCNT improves survival and the number of pronuclei in reconstructed rat embryos after activation. These reconstructed embryos treated before, during, and after SCNT showed significantly higher p34(cdc2) kinase activity involving CSF activity compared to that of the control embryos. On the other hand, p34(cdc2) kinase activity was not recovered in nuclear-transferred oocytes without MG132, which suggested that the enucleation had detrimental effects on the development of reconstructed oocytes. Taken together, MG132 treatment during SCNT increases survival and pronuclear numbers in reconstructed rat embryos via maintenance of high CSF activity. The data suggest that MG132 treatment is indispensable for at least rat SCNT.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"461-8"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0038","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27822329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The differentiation of hepatocyte-like cells from monkey embryonic stem cells.","authors":"Xiaocui Ma,&nbsp;Yuyou Duan,&nbsp;Christine J Jung,&nbsp;Jian Wu,&nbsp;Catherine A VandeVoort,&nbsp;Mark A Zern","doi":"10.1089/clo.2007.0012","DOIUrl":"https://doi.org/10.1089/clo.2007.0012","url":null,"abstract":"<p><p>Embryonic stem cells (ESC) hold great potential for the treatment of liver diseases. Here, we report the differentiation of rhesus macaque ESC along a hepatocyte lineage. The undifferentiated monkey ESC line, ORMES-6, was cultured in an optimal culture condition in an effort to differentiate them into hepatocyte-like cells in vitro. The functional efficacy of the differentiated hepatic cells was evaluated using RT-PCR for the expression of hepatocyte specific genes, and Western blot analysis and immunocytochemistry for hepatic proteins such as alpha-fetoprotein (AFP), albumin and alpha1-antitrypsin (alpha1-AT). Functional assays were performed using the periodic acid schiff (PAS) reaction and ELISA. The final yield of ESC-derived hepatocyte-like cells was measured by flow cytometry for cells that were transduced with a liver-specific lentivirus vector containing the alpha1-AT promoter driving the expression of green fluorescence protein (GFP). The treatment of monkey ESC with an optimal culture condition yielded hepatocyte-like cells that expressed albumin, alpha1-AT, AFP, hepatocyte nuclear factor 3beta, glucose-6-phophatase, and cytochrome P450 genes and proteins as determined by RT-PCR and Western blot analysis. Immunofluorescent staining showed the cells positive for albumin, AFP, and alpha1-AT. PAS staining demonstrated that the differentiated cells showed hepatocyte functional activity. Albumin could be detected in the medium after 20 days of differentiation. Flow cytometry data showed that 6.5 +/- 1.0% of the total differentiated cells were positive for GFP. These results suggest that by using a specific, empirically determined, culture condition, we were able to direct monkey ESC toward a hepatocyte lineage.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"485-93"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2007.0012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27678854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chemical enhancement in embryo development and stem cell derivation from single blastomeres.","authors":"Chanchao Lorthongpanich, Shang-Hsun Yang, Karolina Piotrowska-Nitsche, Rangsun Parnpai, Anthony W S Chan","doi":"10.1089/clo.2008.0035","DOIUrl":"10.1089/clo.2008.0035","url":null,"abstract":"<p><p>Several chemicals targeting the mitogen-activated protein (MAP) kinase signaling pathway, which play an important role in regulating cell growth and differentiation, have shown enhancing effects on the development of the inner cell mass (ICM) and the derivation of ES cells. However, investigation of such chemicals on early embryonic development and the establishment of ES cell lines has not been elucidated. This study was aimed to determine if ACTH, MAP2K1 inhibitor [MAP2K1 (I)], and MAPK14 inhibitor [MAPK14 (I)] could enhance the development of the ICM in preimplantation mouse embryos and blastocyst outgrowths, and the establishment of ES cell lines from blastomeres of early embryos. We have demonstrated that both MAP2K1 (I) and MAPK14 (I) delay early embryo development and inhibit the development of embryos from early blastomeres. On the other hand, ACTH had a positive effect on embryos derived from early blastomeres. As a result, 17 ES cell lines were established. Among these ES cell lines, nine and five ES cell lines were established from single blastomeres of two-cell embryos with and without the supplement of ACTH, respectively. In addition to two-cell isolated blastomeres, three ES cell lines were established from blastomeres of four-cell embryos only with the supplement of ACTH. Our results suggest that ACTH can enhance the derivation of ES cells from single blastomere-derived embryos.</p>","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"10 4","pages":"503-12"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3140851/pdf/clo.2008.0035.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27678856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}