Human CD8 T cells generated in vitro from hematopoietic stem cells are functionally mature

<p>Abstract</p> <p>Background</p> <p>T cell development occurs within the highly specialized thymus. Cytotoxic CD8 T cells are critical in adaptive immunity by targeting virally infected or tumor cells. In this study, we addressed whether functional CD8 T cells can be generated fully <it>in vitro </it>using human umbilical cord blood (UCB) hematopoietic stem cells (HSCs) in coculture with OP9-DL1 cells.</p> <p>Results</p> <p>HSC/OP9-DL1 cocultures supported the differentiation of CD8 T cells, which were TCR/CD3^hiCD27^hiCD1a^negand thus phenotypically resembled mature functional CD8 single positive thymocytes. These <it>in vitro</it>-generated T cells also appeared to be conventional CD8 cells, as they expressed high levels of <it>Eomes </it>and low levels of <it>Plzf</it>, albeit not identical to <it>ex vivo </it>UCB CD8 T cells. Consistent with the phenotypic and molecular characterization, upon TCR-stimulation, <it>in vitro</it>-generated CD8 T cells proliferated, expressed activation markers (MHC-II, CD25, CD38), secreted IFN-γ and expressed Granzyme B, a cytotoxic T-cell effector molecule.</p> <p>Conclusion</p> <p>Taken together, the ability to direct human hematopoietic stem cell or T-progenitor cells towards a mature functional phenotype raises the possibility of establishing cell-based treatments for T-immunodeficiencies by rapidly restoring CD8 effector function, thereby mitigating the risks associated with opportunistic infections.</p

Uterine Compression Sutures as an Effective Treatment for Postpartum Hemorrhage: Case Series

We evaluated the role of uterine compression sutures as a conservative treatment for postpartum hemorrhage (PPH) after failed medical treatment. We retrospectively reviewed the charts of all patients who delivered between 2003 and 2009 at a single tertiary care center and who underwent uterine compression sutures for PPH. Twelve women had uterine compression sutures for PPH. The mean age of the patients was 36.3 ± 5.2 years. The mean gestational age at delivery was 37.7 ± 2.0 weeks, and the average estimated blood loss was 2.1 ± 1.1 L. The mean procedure time to perform the uterine compression sutures was 9.3 ± 2.8 minutes. The success rate of compression sutures was 92% with only one failure resulting in a hysterectomy. Uterine compression sutures are an effective method for the treatment of PPH, thus avoiding hysterectomy and preserving potential fertility

Exosomes from acellular Wharton’s jelly of the human umbilical cord promotes skin wound healing

Abstract Background Compromised wound healing has become a global public health challenge which presents a significant psychological, financial, and emotional burden on patients and physicians. We recently reported that acellular gelatinous Wharton’s jelly of the human umbilical cord enhances skin wound healing in vitro and in vivo in a murine model; however, the key player in the jelly which enhances wound healing is still unknown. Methods We performed mass spectrometry on acellular gelatinous Wharton’s jelly to elucidate the chemical structures of the molecules. Using an ultracentrifugation protocol, we isolated exosomes and treated fibroblasts with these exosomes to assess their proliferation and migration. Mice were subjected to a full-thickness skin biopsy experiment and treated with either control vehicle or vehicle containing exosomes. Isolated exosomes were subjected to further mass spectrometry analysis to determine their cargo. Results Subjecting the acellular gelatinous Wharton’s jelly to proteomics approaches, we detected a large amount of proteins that are characteristic of exosomes. Here, we show that the exosomes isolated from the acellular gelatinous Wharton’s jelly enhance cell viability and cell migration in vitro and enhance skin wound healing in the punch biopsy wound model in mice. Mass spectrometry analysis revealed that exosomes of Wharton’s jelly umbilical cord contain a large amount of alpha-2-macroglobulin, a protein which mimics the effect of acellular gelatinous Wharton’s jelly exosomes on wound healing. Conclusions Exosomes are being enriched in the native niche of the umbilical cord and can enhance wound healing in vivo through their cargo. Exosomes from the acellular gelatinous Wharton’s jelly and the cargo protein alpha-2-macroglobulin have tremendous potential as a noncellular, off-the-shelf therapeutic modality for wound healing

Additional file 3: of Exosomes from acellular Wharton’s jelly of the human umbilical cord promotes skin wound healing

Video S1. Time-lapse imaging showing that exosomes concentrate to the cell periphery and, by 3 h, disappear as depicted by a decrease in red fluorescence. (WMV 32791 kb

Additional file 1: of Exosomes from acellular Wharton’s jelly of the human umbilical cord promotes skin wound healing

Figure S1. Scanning electron microscopy (SEM) images of cord cross-section. SEM images show two zones. Zone 1 displays a section in between the umbilical vein and two arteries, and zone 2 displays a region closer to the umbilical vein. (A–E) Magnification of zone 1. (F–J) Magnification of zone 2. (TIF 10880 kb

Additional file 4: of Exosomes from acellular Wharton’s jelly of the human umbilical cord promotes skin wound healing

Figure S3. Cell viability quantification for keratinocytes. Control medium-treated keratinocytes compared with exosome-treated and α2M (100 ng/ml)-treated keratinocytes. *p < 0.05, ***p < 0.001. N = 6 for control and exosomes; N = 12 for α2M. (TIF 4635 kb

Additional file 2: of Exosomes from acellular Wharton’s jelly of the human umbilical cord promotes skin wound healing

Figure S2. Schematic representation of the exosome isolation protocol. (TIF 3668 kb

Human Wharton’s jelly mesenchymal stem cells promote skin wound healing through paracrine signaling

Abstract Introduction The prevalence of nonhealing wounds is predicted to increase due to the growing aging population. Despite the use of novel skin substitutes and wound dressings, poorly vascularized wound niches impair wound repair. Mesenchymal stem cells (MSCs) have been reported to provide paracrine signals to promote wound healing, but the effect of human Wharton’s jelly-derived MSCs (WJ-MSCs) has not yet been described in human normal skin. The aim of this study is to examine the effects of human WJ-MSC paracrine signaling on normal skin fibroblasts in vitro, and in an in vivo preclinical model. Methods Human WJ-MSCs and normal skin fibroblasts were isolated from donated umbilical cords and normal adult human skin. Fibroblasts were treated with WJ-MSC-conditioned medium (WJ-MSC-CM) or nonconditioned medium. Results Expression of genes involved in re-epithelialization (transforming growth factor-β2), neovascularization (hypoxia-inducible factor-1α) and fibroproliferation (plasminogen activator inhibitor-1) was upregulated in WJ-MSC-CM-treated fibroblasts (P ≤ 0.05). WJ-MSC-CM enhanced normal skin fibroblast proliferation (P ≤ 0.001) and migration (P ≤ 0.05), and promoted wound healing in an excisional full-thickness skin murine model. Conclusions Under our experimental conditions, WJ-MSCs enhanced skin wound healing in an in vivo mouse model

Maternal and Neonatal Morbidity for Women Who Would Be Added to the Diagnosis of GDM Using IADPSG Criteria: A Secondary Analysis of the Hyperglycemia and Adverse Pregnancy Outcome Study

To assess the frequency of adverse outcomes for women who are diagnosed with gestational diabetes mellitus (GDM) by the International Association of the Diabetes and Pregnancy Study Groups (IADPSG) criteria using data from the Hyperglycemia and Adverse Pregnancy Outcome (HAPO) study.This is a secondary analysis from the North American HAPO study centers. Glucose measurements from a 75-g oral glucose tolerance test were used to group participants into three nonoverlapping categories: GDM based on Carpenter-Coustan (CC) criteria (also GDM based on IADPSG criteria), GDM diagnosed based on IADPSG criteria but not CC criteria, and no GDM. Newborn outcomes included birth weight, cord C-peptide, and newborn percentage fat above the 90th percentile; maternal outcomes included primary cesarean delivery and preeclampsia. Outcome frequencies were compared using multiple logistic regression, adjusting for predefined covariates.Among 25,505 HAPO study participants, 6,159 blinded participants from North American centers were included. Of these, 81% had normal glucose testing, 4.2% had GDM based on CC criteria, and 14.3% had GDM based on IADPSG criteria but not CC criteria. Compared with women with no GDM, those diagnosed with GDM based on IADPSG criteria had adjusted odds ratios (95% CIs) for birth weight, cord C-peptide, and newborn percentage fat above the 90th percentile, as well as primary cesarean delivery and preeclampsia, of 1.87 (1.50-2.34), 2.00 (1.54-2.58), 1.73 (1.35-2.23), 1.31 (1.07-1.60), and 1.73 (1.32-2.27), respectively.Women diagnosed with GDM based on IADPSG criteria had higher adverse outcome frequencies compared with women with no GDM. These data underscore the need for research to assess the effect of treatment to improve outcomes in such women