ABSTRACT
The genetic regulatory network controlling early fate choices during human blood cell development are not well understood. We used human pluripotent stem cell reporter lines to track the development of endothelial and haematopoietic populations in an in vitro model of human yolk-sac development. We identified SOX17-CD34+CD43- endothelial cells at day 2 of blast colony development, as a haemangioblast-like branch point from which SOX17-CD34+CD43+ blood cells and SOX17+CD34+CD43- endothelium subsequently arose. Most human blood cell development was dependent on RUNX1. Deletion of RUNX1 only permitted a single wave of yolk sac-like primitive erythropoiesis, but no yolk sac myelopoiesis or aorta-gonad-mesonephros (AGM)-like haematopoiesis. Blocking GFI1 and/or GFI1B activity with a small molecule inhibitor abrogated all blood cell development, even in cell lines with an intact RUNX1 gene. Together, our data define the hierarchical requirements for RUNX1, GFI1 and/or GFI1B during early human haematopoiesis arising from a yolk sac-like SOX17-negative haemogenic endothelial intermediate.
Subject(s)
Blood Cells/metabolism , Core Binding Factor Alpha 2 Subunit/metabolism , DNA-Binding Proteins/metabolism , Endothelium/metabolism , Hematopoiesis , Proto-Oncogene Proteins/metabolism , Repressor Proteins/metabolism , SOXF Transcription Factors/metabolism , Transcription Factors/metabolism , Yolk Sac/metabolism , Blood Cells/cytology , Cell Differentiation , Cell Lineage , Erythroid Cells/cytology , Erythroid Cells/metabolism , Histone Demethylases/antagonists & inhibitors , Histone Demethylases/metabolism , Humans , Models, Biological , Transcription, GeneticABSTRACT
We describe the generation and characterization of 5 human induced pluripotent stem cell (iPSC) lines derived from peripheral blood mononuclear cells (PBMCs) of healthy adult individuals. The PBMCs were reprogrammed using non-integrating Sendai viruses containing the reprogramming factors POU5F1 (OCT4), SOX2, KLF4 and MYC. The iPSC lines exhibited a normal karyotype, expressed pluripotency markers and differentiated into cells representative of the three embryonic germ layers. These iPSC lines can be used as controls in studying disease mechanisms.
Subject(s)
Cell Culture Techniques/methods , Induced Pluripotent Stem Cells/pathology , Leukocytes, Mononuclear/pathology , Adult , Cell Line , Female , Humans , Kruppel-Like Factor 4 , Male , Middle Aged , Young AdultABSTRACT
PURPOSE: The purpose of this study was to determine the common meanings a history of violence has for women out of abusive and violent relationships with an intimate male partner for 5 or more years. SPECIFIC AIMS: To describe the common meanings and shared practices of women who left violent and abusive heterosexual intimate relationships 5 or more years ago, the challenges they face in their current lives, and the resources they use to meet those challenges. An additional aim is to elucidate practical advice they have for others who want to be supportive of the efforts of women recovering from intimate partner violence. METHODS: An interpretive phenomenological approach using Heideggerian hermeneutics was utilized. Approval of the University Social Sciences Institutional Review Board was obtained. Participants were recruited by means of fliers distributed through a domestic violence listserv and through postings in health clinics in western New York. Interviews were recorded and transcribed. A hermeneutic team approach was used for analysis and interpretation of texts. RESULTS: Twenty-one women of various ages, ethnicities, and backgrounds, who self-identified as being out of abusive relationships for 5 or more years, were interviewed. Six themes were identified: developing and maintaining self reliance; negotiating relationships; creating a safe and supportive environment; challenging societal roles and expectations; nurturing the self; and protecting the children. Engendering independence while living with purpose was the constitutive pattern that unified the themes. CONCLUSIONS: Women can successfully establish productive, meaningful lives after violence and will fiercely protect and maintain their independence as they negotiate relationships and developmental challenges throughout their lives. A need for control of their lives and difficulty trusting others remain a lasting legacy of living with a history of violence. CLINICAL RELEVANCE: This is the first study that examines women's lives 5 or more years after leaving violent and abusive relationships. Findings indicate that women can successfully leave violent and abusive relationships, and challenge the widely accepted notion that women do not leave.
