My contemporaries in 20^<th> century.

De novo copy number variations (CNVs) identified in the 1210B2 iPSC-derived NSPCs. (XLSX 39 kb

Bridging the great divide? Making sense of the human rights-CSR relationship in UK multinational companies

Human rights (HR) and corporate social responsibility (CSR) are both fields of knowledge and research that have been shaped by, and examine, the role of multi-national enterprises in society. Whilst scholars have highlighted the overlapping nature of CSR and HR, our understanding of this relationship within business practice remains vague and under-researched. To explore the interface between CSR and HR, this paper presents empirical data from a qualitative study involving 22 international businesses based in the UK. Through an analysis based on sensemaking, the paper examines how and where CSR and HR overlap, contrast and shape one another, and the role that companies’ international operations has on this relationship. The findings reveal a complex and multi-layered relationship between the two, and concludes that in contrast to management theory, companies have bridged the ‘great divide’ in varying degrees most notably in their implementation strategies

Feeder-Free Generation and Long-Term Culture of Human Induced Pluripotent Stem Cells Using Pericellular Matrix of Decidua Derived Mesenchymal Cells

<div><p>Human ES cells (hESCs) and human induced pluripotent stem cells (hiPSCs) are usually generated and maintained on living feeder cells like mouse embryonic fibroblasts or on a cell-free substrate like Matrigel. For clinical applications, a quality-controlled, xenobiotic-free culture system is required to minimize risks from contaminating animal-derived pathogens and immunogens. We previously reported that the pericellular matrix of decidua-derived mesenchymal cells (PCM-DM) is an ideal human-derived substrate on which to maintain hiPSCs/hESCs. In this study, we examined whether PCM-DM could be used for the generation and long-term stable maintenance of hiPSCs. Decidua-derived mesenchymal cells (DMCs) were reprogrammed by the retroviral transduction of four factors (OCT4, SOX2, KLF4, c-MYC) and cultured on PCM-DM. The established hiPSC clones expressed alkaline phosphatase, hESC-specific genes and cell-surface markers, and differentiated into three germ layers in vitro and in vivo. At over 20 passages, the hiPSCs cultured on PCM-DM held the same cellular properties with genome integrity as those at early passages. Global gene expression analysis showed that the GDF3, FGF4, UTF1, and XIST expression levels varied during culture, and GATA6 was highly expressed under our culture conditions; however, these gene expressions did not affect the cells’ pluripotency. PCM-DM can be conveniently prepared from DMCs, which have a high proliferative potential. Our findings indicate that PCM-DM is a versatile and practical human-derived substrate that can be used for the feeder-cell-free generation and long-term stable maintenance of hiPSCs.</p> </div

In vivo differentiation of hiPSCs generated on PCM-DM at early and late passages.

<p>hiPSC-PCMDM-induced teratomas were excised from mice and processed for H&E staining. Clone 1, iPS-DMC72-PCMDM01; Clone 2, iPS-DMC72-PCMDM02. Early, passage 12; Late, passage 23; Scale bar = 100 µm.</p

Increased GATA6 expression in 201B7 on PCM-DM with StemPro medium.

<p>A) Quantitative RT-PCR analysis of OCT4 and GATA6 for 201B7 cultured on PCM-DM with MEF-CM or StemPro medium. “+number” indicates the passage number after reseeding on PCM-DM. Statistical significances are determined by Scheffe’s test after two-way ANOVA. Results of comparisons among groups of medium within each passage are shown (*, P<0.01). B) Morphology of 201B7 cultured on PCM-DM with MEF-CM or StemPro medium. Scale bar = 500 µm.</p

In vitro characterization of hiPSCs generated on PCM-DM.

<p>A) Quantitative RT-PCR analysis for the mRNA copy number of four transgenes (OCT4, SOX2, KLF4, c-MYC). All the transgenes were silenced in two hiPSC-PCMDM clones. Data are presented as the mean ± SD. Clone 1, iPS-DMC72-PCMDM01; Clone 2, iPS-DMC72-PCMDM02; Early, passage 8; Late, passage 30; *: not detected. B) Quantitative RT-PCR analysis for hESC marker gene (OCT4, SOX2, KLF4, c-MYC, NANOG) expression at early (passage 8) and late (passage 30) culture times compared with hESCs (clone KhES1). Data are presented as the mean ± SD. C) Immunocytochemistry for NANOG (red) and OCT4 (green) expression in hiPSC-PCMDM clones. Clone 1, Early (passage 10), Late (passage 22); Clone 2, Early (passage 11), Late (passage 25). Scale bar = 200 µm. D) Flow cytometry analysis for hESC-specific surface antigens (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81) at early (passage 10) and late (passage 30) culture times in comparison with parental DMCs and hESCs (KhES1). Clone 1, iPS-DMC72-PCMDM01; Clone2, iPS-DMC72-PCMDM02.</p

In vitro differentiation of hiPSCs generated on PCM-DM at early and late passages.

<p>A) Embryoid bodies (EBs) on day 8, derived from hiPSCs generated on PCM-DM at early (Clone 1, passage 11; Clone 2, passage 12) and late (Clone 1, passage 22; Clone 2, passage 21) culture times. Scale bar = 200 µm. B) Quantitative RT-PCR analysis. Left: Expression levels of undifferentiated genes in EBs relative to the hiPSCs before differentiation. Right: Expression levels of lineage-specific genes in EBs relative to differentiated 201B7. Data are presented as the mean ± SD. C) Immunocytochemical staining of differentiated cells by culturing EBs on gelatin-coated chamber slides with DMEM containing 10% FBS for 1 week. Clone 1, iPS-DMC72-PCMDM01; Clone 2, iPS-DMC72-PCMDM02. Scale bar = 50 µm.</p

Generation of hiPSCs from DMCs with non-conditioned hESC medium on PCM-DM.

<p>A) Morphology and Alkaline phosphatase (ALP) staining of iPS-DMC75-PCMDM. P, passage number. Scale bar = 500 µm. B). Quantitative RT-PCR analysis for the mRNA copy number of four transgenes (OCT4, SOX2, KLF4, c-MYC). All the transgenes were silenced in iPS-DMC75-PCMDM. Data are presented as the mean ± SD. *: not detected. C) Immunocytochemistry for NANOG (red) and OCT4 (green) expression in iPS-DMC75-PCMDM. Scale bar = 200 µm. D) Flow cytometry analysis for hESC-specific surface antigens (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81) at passage 11. E) Global gene expression analysis of iPS-DMC75-PCMDM (passage 12), early and late passages of clones 1 and 2, KhES1, and 201B7. Scatter plots and Pearson’s coefficient are shown. The diagonal lines indicate 3-fold changes in gene expression levels. Plus (“+”) symbols indicate stem-cell marker genes suggested by the International Stem Cell Initiative <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055226#pone.0055226-Adewumi1" target="_blank">[31]</a>, and such genes outside the 3-fold change lines are shown in red text. Clone 1, iPS-DMC72-PCMDM01; Clone 2, iPS-DMC72-PCMDM02. F) Embryoid bodies (EBs) on day 8 derived from iPS-DMC75-PCMDM (passage 12). Scale bar = 500 µm. G) Quantitative RT-PCR analysis. Left, Expression levels of genes for the undifferentiated state in EBs relative to hiPSCs before differentiation. Right, Expression levels of lineage-specific genes in EBs relative to differentiated 201B7. Data are presented as the mean ± SD.</p

Global gene expression analysis.

<p>A) Global gene expression of the hiPSC-PCMDM clones, KhES1, and 201B7 by microarray analysis. Scatter plots and Pearson’s coefficient are shown. The diagonal lines indicate 3-fold changes in gene expression levels. Plus (“+”) symbols indicate stem-cell marker genes suggested by the International Stem Cell Initiative <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055226#pone.0055226-Adewumi1" target="_blank">[31]</a>, and such genes outside the 3-fold change lines are shown in red text. Early, passage 8; Late, passage 30. Clone 1, iPS-DMC72-PCMDM01; Clone 2, iPS-DMC72-PCMDM02. B) Quantitative RT-PCR analysis of the XIST, GDF3, FGF4, UTF1, and GATA6 genes. Data are presented as the mean ± SD. Early, passage 8; Late, passage 30. Statistical differences between early and late passage group are determined by unpaired Student’s t-test (*, P<0.01). C) Hierarchical cluster analysis between parental DMCs, KhES1 cells, 201B7, and the two hiPSC-PCMDM clones at early (passage 8) and late (passage 30) passages.</p

Karyotype and promoter methylation analyses of hiPSCs generated on PCM-DM.

<p>A) G-band staining of the two hiPSC-PCMDM clones showing a normal female karyotype (46, XX) in both clones at early (Clone 1, passage 12; Clone 2, passage 12) and late (Clone 1, passage 23; Clone 2, passage 24) passages. B) Methylation states of the OCT4 and NANOG promoter of the two hiPSC-PCMDM clones (passage 29) using bisulphate sequencing. Numbers indicate the position from the transcription start site. Open squares indicate unmethylated and filled squares indicate methylated CpG dinucleotides. Clone 1, iPS-DMC72-PCMDM01; Clone 2, iPS-DMC72-PCMDM02.</p