Cloning Stem Cells最新文献_第6页

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0052

Josef Fulka, Pasqualino Loi, Grazyna Ptak, Helena Fulka, Justin St John

引用次数: 16

Gene expression profile of multipotent mesenchymal stromal cells: Identification of pathways common to TGFbeta3/BMP2-induced chondrogenesis. 多能间充质间质细胞的基因表达谱:TGFbeta3/ bmp2诱导软骨形成的共同途径的鉴定。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0070

Dominique Mrugala, Nadège Dossat, Jochen Ringe, Bruno Delorme, Amandine Coffy, Claire Bony, Pierre Charbord, Thomas Häupl, Jean-Pierre Daures, Danièle Noël, Christian Jorgensen

{"title":"Gene expression profile of multipotent mesenchymal stromal cells: Identification of pathways common to TGFbeta3/BMP2-induced chondrogenesis.","authors":"Dominique Mrugala, Nadège Dossat, Jochen Ringe, Bruno Delorme, Amandine Coffy, Claire Bony, Pierre Charbord, Thomas Häupl, Jean-Pierre Daures, Danièle Noël, Christian Jorgensen","doi":"10.1089/clo.2008.0070","DOIUrl":"https://doi.org/10.1089/clo.2008.0070","url":null,"abstract":"Multipotent mesenchymal stromal cells (MSC) display a high potential for the development of novel treatment strategies for cartilage repair. However, the pathways involved in their differentiation to functional non hypertrophic chondrocytes remain largely unknown, despite the work on embryologic development and the identification of key growth factors including TGFbeta, Hh, Wnt and FGF. In this study, we asked if we could identify specific biological networks common to the growth factors used (TGFbeta3 or BMP-2). To address this question, we used DNA microarrays and performed large-scale expression profiling of MSC at different time points during their chondrogenic differentiation. By comparing these data with those obtained during the differentiation of MSC into osteoblasts and adipocytes, we identified 318 genes specific for chondrogenesis and developed a new algorithm to classify the genes according to their kinetic profile. We distributed the selected genes in five classes according to their kinetic of expression. We could reconstruct three phases characterized by functional pathways. The first phase corresponds to cell attachment and apoptosis induction; the second phase is characterized by a proliferation/differentiation step, and the third phase is characterized by a differentiation/hypertrophy pathway. Indeed, these data propose new pathways to understand the complexity of MSC differentiation to chondrocytes.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"61-76"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0070","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27968431","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}

引用次数: 52

Tracing the stemness of porcine skin-derived progenitors (pSKP) back to specific marker gene expression. 猪皮肤源性祖细胞(pSKP)的干性溯源至特定的标记基因表达。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0071

Mingtao Zhao, S Clay Isom, Hui Lin, Yanhong Hao, Yong Zhang, Jianguo Zhao, Jeffrey J Whyte, Kyle B Dobbs, Randall S Prather

{"title":"Tracing the stemness of porcine skin-derived progenitors (pSKP) back to specific marker gene expression.","authors":"Mingtao Zhao, S Clay Isom, Hui Lin, Yanhong Hao, Yong Zhang, Jianguo Zhao, Jeffrey J Whyte, Kyle B Dobbs, Randall S Prather","doi":"10.1089/clo.2008.0071","DOIUrl":"https://doi.org/10.1089/clo.2008.0071","url":null,"abstract":"Multipotent skin-derived progenitors (SKP) can produce both neural and mesodermal progeny in vitro, sharing the characteristics of embryonic neural crest stem cells. However, the molecular basis for the property of multiple lineage potential and neural crest origin of SKPs is still elusive. Here we report the cooperative expression of pluripotency related genes (POU5F1, SOX2, NANOG, STAT3) and neural crest marker genes (p75NTR, TWIST1, PAX3, SNAI2, SOX9, SOX10) in GFP-transgenic porcine skin-derived progenitors (pSKP). The proportion of cells positive for POU5F1, nestin, fibronectin, and vimentin were 12.3%, 15.1%, 67.9% and 53.7%, showing the heterogeneity of pSKP spheres. Moreover, pSKP cells can generate both neural (neurons and glia) and mesodermal cell types (smooth muscle cells and adipocytes) in vitro, indicating the multiple lineage potency. Four transcription factors (POU5F1, SNAI2, SOX9, and PAX3) were identified that were sensitive to mitogen (FBS) and/or growth factors (EGF and bFGF). We infer that POU5F1, SNAI2, SOX9, and PAX3 may be the key players for maintaining the neural crest derived multipotency of SKP cells in vitro. This study has provided new insight into the molecular mechanism of stemness for somatic-derived stem cells at the level of transcriptional regulation.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"111-22"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0071","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27992695","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}

引用次数: 40

Improvement of embryonic stem cell line derivation efficiency with novel medium, glucose concentration, and epigenetic modifications. 利用新型培养基、葡萄糖浓度和表观遗传修饰提高胚胎干细胞系衍生效率。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0053

Chul Kim, Tomokazu Amano, Joonghoon Park, Mark G Carter, Xiuchun Tian, Xiangzhong Yang

{"title":"Improvement of embryonic stem cell line derivation efficiency with novel medium, glucose concentration, and epigenetic modifications.","authors":"Chul Kim, Tomokazu Amano, Joonghoon Park, Mark G Carter, Xiuchun Tian, Xiangzhong Yang","doi":"10.1089/clo.2008.0053","DOIUrl":"https://doi.org/10.1089/clo.2008.0053","url":null,"abstract":"Although the first mouse embryonic stem (ES) cell lines were derived 2 decades ago, and standard protocols for ES cell derivation are widely used today, the technical difficulty of these protocols still pose a challenge for many investigators attempting to produce large numbers of ES cell lines, and are limited to only a few mouse strains. Recently, glucose concentration was shown to have a significant effect on the efficiency of ES cell derivation, but the mechanism(s) mediating this effect are still the subject of debate. In this report, we investigated the effect of glucose concentration on ES cell derivation efficiency from blastocysts in the context of a new medium, Minimum Essential Medium alpha (MEMalpha). Furthermore, we propose novel methods to improve mouse ES cell derivation efficiency using in vitro epigenetic modifications during early passages, combined with detection of Oct4-expressing cells. Based on the results reported here, modified MEMalpha containing high glucose improves the efficiency of ES cell derivation remarkably, compared with Knockout Dulbecco's-Modified Eagle Media (KDMEM). Epigenetic modifications are able to improve the efficiency even further.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"89-100"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0053","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27992696","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}

引用次数: 19

Human embryos derived by somatic cell nuclear transfer using an alternative enucleation approach. 利用另一种去核方法进行体细胞核移植获得的人类胚胎。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0041

Jianyuan Li, Xuexia Liu, Haiyan Wang, Shouxin Zhang, Fujun Liu, Xuebo Wang, Yanwei Wang

{"title":"Human embryos derived by somatic cell nuclear transfer using an alternative enucleation approach.","authors":"Jianyuan Li, Xuexia Liu, Haiyan Wang, Shouxin Zhang, Fujun Liu, Xuebo Wang, Yanwei Wang","doi":"10.1089/clo.2008.0041","DOIUrl":"https://doi.org/10.1089/clo.2008.0041","url":null,"abstract":"Somatic cell nuclear transfer (SCNT) was used to generate patient-specific embryonic stem cells (ESCs) from blastocysts cloned by nuclear transfer (ntESCs). In this study, a total of 135 oocytes were obtained from 12 healthy donors (30-35 years). Human oocytes, obtained within 2 h following transvaginal aspiration, were enucleated using a Spindle Imaging System to position the spindle and chromosomes that lay on the metaphase plate, and a Zona Infrared Laser Optical System was used to open a single hole in the zona pellucida at the ~ 2 o'clock position. Human fibroblasts and lymphocytes were used to construct SCNT embryos. Nearly half (26 of 58) of the oocytes were fused after electrofusion and embryo development rates were 96.2% (two-cell, 25 of 26), 92.3% (four-cell, 24 of 26), 61.5% (eight-cell, 16 of 26), 34.6% (16-cell, 9 of 26), 26.9% (morula, 7 of 26), and 19.2% (blastocyst, 5 of 26), respectively, following incubation in improved G-series sequential medium. One cloned blastocyst was used for STR-DNA identification and genetic polymorphism analysis of mtDNA, and STR-DNA analysis of all cloned blastocysts indicated they were derived from SCNT. Quantitative analysis showed that mtDNA of cloned embryos reflected the change tendency of those observed in human IVF embryos. Our research provides an alternative enucleation approach for producing human SCNT-derived blastocysts, and may aid in providing a new method for human therapeutic cloning.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"39-50"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0041","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27968434","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}

引用次数: 36

Direct Differentiation of Human Embryonic Stem Cells to Hepatocyte-like Cells Exhibiting Functional Activities (vol 9, pg 51, 2007) 人类胚胎干细胞向具有功能活性的肝细胞样细胞的直接分化(vol 9, pg 51, 2007)

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/CLO.2008.00C1

J. Lebkowski, W. Cui, D. Hay

引用次数: 0

Porcine skin-derived stem cells can serve as donor cells for nuclear transfer. 猪皮肤源性干细胞可作为核移植供体细胞。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0063

Yanhong Hao, David Wax, Zhisheng Zhong, Clifton Murphy, Jason W Ross, August Rieke, Melissa Samuel, Lee Spate, Paul Dyce, Julang Li, Peter Sutovsky, Randall S Prather

{"title":"Porcine skin-derived stem cells can serve as donor cells for nuclear transfer.","authors":"Yanhong Hao, David Wax, Zhisheng Zhong, Clifton Murphy, Jason W Ross, August Rieke, Melissa Samuel, Lee Spate, Paul Dyce, Julang Li, Peter Sutovsky, Randall S Prather","doi":"10.1089/clo.2008.0063","DOIUrl":"https://doi.org/10.1089/clo.2008.0063","url":null,"abstract":"Although transgenic animal production through somatic cell nuclear transfer (SCNT) has been successful, the process is still inefficient. One major limitation is the use of somatic donor cells that have a finite life span. Identification and isolation of a cell type capable of rapid proliferation while possessing immortal or prolonged life span in culture and is capable of being genetically modified would be very valuable for utilization in the production of genetically modified pigs. Here we report the birth of live piglets after cloning by using porcine skin-derived stem cells (SSC) as a donor cell type. In the present study, cell cycle analysis indicates that the porcine SSC proliferate rapidly in vitro. The porcine SSC are capable of producing live offspring and can be genetically modified with positive selection. Utilization of porcine SSC may prove to be an excellent cell type for genetic modification followed by nuclear transfer for the production of transgenic pigs.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"101-10"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0063","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27992721","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}

引用次数: 28

Generation of domestic transgenic cloned kittens using lentivirus vectors. 用慢病毒载体培育国内转基因克隆小猫。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0054

Martha C Gómez, Charles Earle Pope, Robert H Kutner, David M Ricks, Leslie A Lyons, Mark T Ruhe, Cherie Dumas, Justine Lyons, Betsy L Dresser, Jakob Reiser

{"title":"Generation of domestic transgenic cloned kittens using lentivirus vectors.","authors":"Martha C Gómez, Charles Earle Pope, Robert H Kutner, David M Ricks, Leslie A Lyons, Mark T Ruhe, Cherie Dumas, Justine Lyons, Betsy L Dresser, Jakob Reiser","doi":"10.1089/clo.2008.0054","DOIUrl":"https://doi.org/10.1089/clo.2008.0054","url":null,"abstract":"The efficient use of somatic cell nuclear transfer (SCNT), in conjunction with genetic modification of donor cells provides a general means to add or inactivate genes in mammals. This strategy has substantially improved the efficacy of producing genetically identical animals carrying mutant genes corresponding to specific human disorders. Lentiviral (LV) vectors have been shown to be well suited for introducing transgenes into cells to be used as donor nuclei for SCNT. In the present study, we established an LV vector-based transgene delivery approach for producing live transgenic domestic cats by SCNT. We have demonstrated that cat fetal fibroblasts can be transduced with EGFP-encoding LV vectors bearing various promoters including the human cytomegalovirus immediate early (hCMV-IE) promoter, the human translation elongation factor 1alpha (hEF-1alpha) promoter and the human ubiquitin C (hUbC) promoter. Among the promoters tested, embryos reconstructed with donor cells transduced with a LV-vector bearing the hUbC promoter displayed sustained transgene expression at the blastocyst stage while embryos reconstructed with LV vector-transduced cells containing hCMV-IE-EGFP or hEF-1alpha-EGFP cassettes did not. After transfer of 291 transgenic cloned embryos into the oviducts of eight recipient domestic cats (mean =36.5 +/- 10.1), three (37.5%) were diagnosed to be pregnant, and a total of six embryos (2.1%) implanted. One live male offspring was delivered by Cesarean section on day 64 of gestation, and two kittens were born dead after premature delivery on day 55. In summary, we report the birth of transgenic cloned kittens produced by LV vector-mediated transduction of donor cells and confirm that cloned kittens express the EGFP reporter transgene in all body tissues.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"167-76"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27973763","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}

引用次数: 24

Cells extract from fetal liver promotes the hematopoietic differentiation of human embryonic stem cells. 胎肝细胞提取物促进人胚胎干细胞的造血分化。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0049

Yu-Xiao Liu, Lei Ji, Wen Yue, Zhi-Feng Yan, Jing Wang, Jia-Fei Xi, Rui Zhang, Xue Nan, Ci-Xian Bai, Lin Chen, Yun-Fang Wang, Xue-Tao Pei

{"title":"Cells extract from fetal liver promotes the hematopoietic differentiation of human embryonic stem cells.","authors":"Yu-Xiao Liu, Lei Ji, Wen Yue, Zhi-Feng Yan, Jing Wang, Jia-Fei Xi, Rui Zhang, Xue Nan, Ci-Xian Bai, Lin Chen, Yun-Fang Wang, Xue-Tao Pei","doi":"10.1089/clo.2008.0049","DOIUrl":"https://doi.org/10.1089/clo.2008.0049","url":null,"abstract":"Here, we have now developed a new inducing system to promote the differentiation of human stem cells (hESCs) toward hematopoietic lineages by the treatment with cells extract of human fetal liver tissue (hFLT). The embryoid bodies (EBs) obtained from human H1 embryonic stem cells were exposed to buffer, hFLT cells extract, heated hFLT cell extract, and cell extract of human liver cells lines-LO2. Then, the feature of EBs in different groups was characterized by real-time RT-PCR and colony-forming assays. The results showed the treatment by hFLT cells extract could activate the hematopoietic genes expression and improve the capacity for hematopoietic progenitor development of hEBs. After that, we cocultured hFLT extract treated hEBs on the hFLSCs (human fetal liver stromal cells) feeder to differentiate them into hematopoietic cells. As a control, untreated hEBs were cocultured on hFLSCs feeder with cytokines. The feature of induced cells from hEBs was characterized by flow cytometry, Wright-Giemsa staining, and colony-forming assays. The results demonstrated that hFLT cells extract was capable of inducing hEBs into hematopoietic cells and combing it with hFLSCs feeder could largely promote hematopoietic differentiation of hESCs. This method may supply a new way to substitute the cytokines required in hematopoietic induction of hESCs.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"51-60"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27987603","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}

引用次数: 11

Antimitotic treatments for chemically assisted oocyte enucleation in nuclear transfer procedures. 核移植过程中化学辅助卵母细胞去核的抗有丝分裂处理。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0031

Nuno Costa-Borges, Maria Teresa Paramio, Gloria Calderón, Josep Santaló, Elena Ibáñez

{"title":"Antimitotic treatments for chemically assisted oocyte enucleation in nuclear transfer procedures.","authors":"Nuno Costa-Borges, Maria Teresa Paramio, Gloria Calderón, Josep Santaló, Elena Ibáñez","doi":"10.1089/clo.2008.0031","DOIUrl":"https://doi.org/10.1089/clo.2008.0031","url":null,"abstract":"Chemically assisted enucleation has been successfully applied to porcine and bovine oocytes to prepare recipient cytoplasts for nuclear transfer procedures. In this study, the antimitotic drugs demecolcine, nocodazole, and vinblastine were first assessed for their ability to induce the formation of cortical membrane protrusions in mouse, goat, and human oocytes. While only 2% of the treated human oocytes were able to form a protrusion, high rates of protrusion formation were obtained both in mouse (84%) and goat oocytes (92%), once the treatment was optimized for each species. None of the antimitotics applied was superior to the others in terms of protrusion formation, but mouse oocytes treated with vinblastine were unable to restore normal spindle morphology after drug removal and their in vitro development after parthenogenetic activation was severely compromised, rendering this antimitotic useless for chemically assisted enucleation approaches. Aspiration of the protrusions in mouse oocytes treated with demecolcine or nocodazole yielded 90% of successfully enucleated oocytes and allowed the extraction of a smaller amount of cytoplasm than with mechanical enucleation, but both enucleation methods resulted in the depletion of spindle-associated gamma-tubulin from the prepared cytoplasts. Treatment of mouse oocytes with demecolcine or nocodazole had no effect on their in vitro development after parthenogenetic activation, or on their ability to repolymerize a new spindle after the removal of the drug or the reconstruction of the treated cytoplasts with a somatic nucleus. Therefore, demecolcine- and nocodazole-assisted enucleation appears as an efficient alternative to mechanical enucleation, which can simplify nuclear transfer procedures.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"153-66"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0031","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27992698","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}

引用次数: 16