Cord Blood Stem Cells Research Articles

Evaluation of Unrestricted Somatic Stem Cells Therapy in an Ex-Vivo and in-Vivo Acquired Severe Aplastic Anemia Model

Background: In acquired severe aplastic anemia (aSAA), oligoclonal expansion of dysregulated CD8+ cytotoxic T cells, abnormal function of CD4+ T helper cells, along with elevated production of IFN-γ and TNF-α have been associated with the apoptosis of hematopoietic stem and progenitor cells (HSPC) (Young, N Engl J Med, 2018). Liao and Cairo et al (Cell Transplantation, 2014) have demonstrated that cord blood-derived unrestricted somatic stem cells (USSCs) could significantly reverse the abnormal/inverted ratio of CD4:CD8 T cells, increases the number of FOXP3A positive Treg cells and decrease the level of IFN-γ in an animal model of recessive dystrophic epidermolysis bullosa (RDEB). Alternative therapies are in great need for patients with aSAA as immunosuppressive therapy (IST) overall responses are sub-optimal. Objectives:To determine the effect of USSCs on HSPC survival, differentiation and phosphorylation pathways in an ex-vivo culture of aSAA with human CD34+ cells.To determine the effect of USSCs administration on blood cell count recovery, bone marrow (BM) cellularity and survival in an in-vivo inflammatory cytokine mediated mice model of aSAA. Design/Methods: Human HSPC were cultured with SCF, FLT3 and human TPO. HSPCs alone, with and without IFN-γ and TNF-α served as controls. USSCs (transwell and direct) were added to the culture. HSPC were harvested and assayed for their survival at day 7 and 14. Annexin-V+/PI+ were referred to as late apoptotic cells. Each experimental condition was set up in triplicate. The cells were cultured at 37°C with 5% CO2. Multi-lineage differentiation capacity was assessed with a selective colony forming units (CFU) assay and compared between experimental groups. Signaling pathways were determined using phosphor-flow cytometric analysis of pSTAT1. For the immune mediated mouse model of aSAA, 8-week-old C57BL/6 mice were irradiated with 6.5Gy TBI. Spleen lymphocytes were collected from FVB donors, and injected 4 hours post-radiation into B6 recipient mice via the retro-orbital vein, at 1-1.5 × 106 cells/recipient in 100μL PBS. Each experimental group (PBS control and USSCs) included 20 mice per group. Mice were continuously monitored for their survival. Peripheral blood (PB) was analyzed using a VETSCAN® HM5 Analyzer. Mean ± SEM of the blood counts was compared between experimental groups. Mice femur bones were extracted and cleaned of soft tissue, fixed and sectioned. Paraffin sections were stained with Hematoxylin and Eosin (H&E) to assess morphology. Results: The myelosuppressive effect of IFN-γ and TNF-α was significantly rescued (4-fold increase) by the addition of USSCs (transwell) at day 7. After 28 days direct co-culture with USSCs increased CD34+ survival as well (p<0.001) from 5 to 35 cells/µl. A significantly higher percent of viable cells (73% versus 63%) was observed among the USSCs treated cells (P<0.05). Phosphor-flow cytometric analysis demonstrated an almost 50% decrease in pSTAT1 in the presence of USSCs (p<0.05). Based on CFU analysis, CD34+ cells cultured in the presence of USSCs were able to produce more CFU (52, SD±3.7) at day 7 (p<0.01) compared to IFN-γ and TNF-α treated cells (16, SD±2.8). Also, USSCs were associated with more BFU (p<0.05) at day 14. In our immune mediated aplastic anemia mice model, treatment with USSCs was associated with substantial prolonged mice survival (p=0.02) (Figure 1a). Evaluation of PB counts demonstrated increased hemoglobin levels at day 14 in the USSCs treated mice (p<0.05) 5.6g/dL (SD±0.6), compared to PBS treated mice 2.1g/dL (SD±0.1). BM evaluation of USSCs treated mice versus PBS control displayed higher cellularity in the USSCs treated sections (Figure 1b). Conclusions: Cellular therapy, such as USSCs, may be instrumental in the treatment of aSAA, specifically by increasing BFU-E and eventually hemoglobin levels. Future studies will need to be done to investigate the exact mechanisms of action and the clinical efficacy of this novel cellular therapy. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Epidemiology, Risk Factors and Outcome of Bacterial, Fungal, and Viral Infections in Pediatric Allogeneic and Autologous Hematopoietic Stem Cell Transplantation Recipients

Objectives Infection remains a major cause of morbidity and mortality after hematopoietic stem cell transplant (HSCT). With recent changes in transplant practices, such as the increasing use of alternative donors and T-cell depletion, how various risk factors interplay and affect the timeline of infections have not been well elucidated in children. While the epidemiology and risk factors for viral infections post-HSCT were well described for cytomegalovirus (CMV) and Epstein-Barr virus (EBV), the corresponding information on human herpes virus 6 (HHV-6), adenovirus (ADV) and BK virus (BKV) infection in children is limited. This study aims to review comprehensively the epidemiology, risk factors and outcome of bacterial, viral and fungal infections after contemporary HSCT to inform future infection prophylaxis strategies. Method We retrospectively reviewed the first 100 consecutive HSCT for malignant and non-malignant diseases from April 2019 to October 2021 in the only pediatric HSCT center in Hong Kong. The incidences were recorded and risk factors were analyzed for bacterial, viral and fungal infections and infection-related mortality. Results The vast majority of the allogeneic transplant recipients (69/74, 93.2%) had infections after HSCT, amongst which bacterial, viral and fungal infection rate was 35.1%, 90.5% and 9.5% respectively. In contrast, only 30.8% (8/26) of autologous transplant recipients had infections, and their respective rate of bacterial, viral and fungal infection was 19.2%, 15.4% and 3.8%. HHV-6 and BKV typically occurred early after HSCT, ADV and varicella zoster virus (VZV) thereafter, and CMV and EBV throughout the entire 2.5-year observation period. The cumulative incidences of HHV-6 and BKV were significantly higher in patients receiving umbilical cord blood (UCB) or haploidentical stem cell transplant (p = 0.0001; p <0.0001, respectively) when compared with matched sibling (MSD) or matched unrelated donor (MUD) transplant (Figure 1 and 2). Ex vivo T-cell depletion was a general risk factor for viral infection with HHV-6 (HR = 3.03), BKV (HR = 3.36), CMV (HR = 4.45) and EBV (HR = 7.15); all p < 0.02. Cancer in second- compared with first-complete remission was a risk factor for bacterial infection in multivariate analyses (OR = 6.0, 95% CI = 1.12-32.2, p = 0.037). Patients with gut graft-versus-host disease were at risk for fungal infections (OR = 12.3, 95% CI = 1.33-114.4, p = 0.027). The infection-related mortality (IRM) rate was 10%, which occurred only in allogeneic HSCT patients with hematological malignancies receiving cord blood (n = 4) or haploidentical HSCT (n = 6). History of bacterial infection after HSCT was associated with an increased risk of IRM (HR = 4.13, 95% CI = 1.05-16.3, p = 0.042). Conclusion Not all HSCT recipients are of equal risk to infections and IRM. Our findings in pediatric patients after contemporary HSCT support both time-dependent and risk-adapted measures against infective complications to improve transplant outcome. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The Investigation of Antitumor Effect of Humanized Bispecific BCMA/CD38 CAR-T Cells on Multiple Myeloma

Background Multiple myeloma (MM) is an incurable disease that will eventually deteriorate. Chimeric antigen receptor (CAR) T cell therapy has been widely used in the treatment of hematological malignancy and showed a promising effect. BCMA (CD269) is exclusively expressed on the MM cells, making it an ideal target for immunotherapy. CD38 is highly expressed on the surface of MM cells, but is also expressed in immune cells and a minority of hematopoietic stem/ progenitor cells. Humanization reshaped antibody may reduce the potential risk of immunogenicity associated with the animal-derived antibodies. In this study, we constructed humanized BCMA CAR-T, CD38 CAR-T and bispecific BCMA/CD38 CAR-T, and explored their killing efficiency towards MM cells, providing the basis for clinical application. Methods Mouse anti-human BCMA and CD38 single chain fragment variable (scFv) sequences were obtained from previously established monoclonal antibody hybridoma cells (69G8 and HIT2) in our institution, respectively. The sequences were then humanized. Single-target CAR (BCMA CAR, CD38 CAR) and dual-target BCMA/CD38 CAR (LoopBC38 CAR) plasmids were constructed. Phenotype of CAR-T cells was detected. CAR-T cells and tumor cell lines were co-cultured in vitro, followed by the detection of the percentage of residual tumor cells, the degranulation and cytokine release level of T cells. Bone marrow mononuclear cells (BMMNC) from MM patients were co-cultured with CAR-T cells. MM cells were intravenously injected into immunodeficient mice, followed by CAR-T cell therapy. After co-culturing with peripheral blood mononuclear cells (PBMC) from healthy donors and CD34+ umbilical cord blood (UCB) stem cells, the proportion, cell subsets of residual cells and colony formation ability were assessed to verify the safety of CAR-T cells. Results The proliferation ability of CD38 CAR-T and LoopBC38 CAR-T was weaker than that of VEC-T and BCMA CAR-T. The residual H929 cells (BCMA+CD38+) after 48h cocultured with VEC-T and LoopBC38 CAR-T at E:T ratio of 1:1 was (20.82±9.773) % vs. (0±0) % (p<0.0001). After 48h at E:T ratio of 1:1, residual K562-BCMA cells (BCMA+CD38-) cocultured with CD38 CAR-T were (49.53±9.773) %, while that of LoopBC38 CAR-T were (8.49±4,766) % (p<0.0001). After 48h at E:T ratio of 1:4, residual Raji cells (BCMA-CD38+) were (62.69±22.2) % in BCMA CAR-T and (19.81±12.22) % in LoopBC38 group (p<0.01). The secretion level of cytokines such as IFN-γ, TNF-α, IL-2 was significantly higher in CAR-T cells than that of VEC-T cells. After co-cultured with H929, the IL-6 secretion level of LoopBC38 CAR-T cells was (7.28±0.4558) %, which was lower than that of CD38 CAR-T cells [(259.4±3.039) %, p<0.0001]. When cocultured with BMMNC derived from MM patients, BCMA CAR-T could kill BCMA+ cells, CD38 CAR T could kill CD38+ cells, while LoopBC38 CAR-T has both effects. In H929 xenograft NSG mice, the median survival time of MM mice was 35 days for VEC-T, 82 days for CD38 CAR-T. Until now, more than 50% mice are alive in BCMA CAR-T group and LoopBC38 CAR-T group, so they didn't reach the median survival time yet. LoopBC38 CAR-T did not significantly kill PBMC form healthy donors, with the residual PBMC cells after 24h co-culturing was (48.26±1.708) % in VEC-T and (49.0±2.517%) in LoopBC38 CAR-T (p>0.05), but had killing efficiency towards mononuclear-macrophages and NK cell. A slight killing effect on UCB CD34+ cells was observed in LoopBC38 CAR-T. The residual CD34+ cells in LoopBC38 CAR-T were (29.47±4.077) %, while that of VEC-T was (48.5±11.88) % (p>0.05). The number of CFU-GEMM in LoopBC38 CAR-T was (3.0±1.512) %, which was lower than that of VEC-T [(4.667±3.386) %, (p<0.001)]. Conclusion All CAR-T cells showed strong killing efficiency toward MM cells both in vitro and in vivo. Humanized BCMA CAR-T and CD38 CAR-T can specifically kill multiple myeloma cell lines and primary cells of patients. Bispecific CAR-T was able to target both antigens. CD38 CAR-T and bispecific CAR-T cells showed self-killing ability and altered their phenotypes. They can kill NK cells, mononuclear-macrophages in peripheral blood of healthy donor and have certain effects on the formation of CFU-GEMM, which may be a safety problem that requires further improvement for the application of CD38 CAR-T and bispecific BCMA/CD38 CAR-T cells.

Cord blood stem cell-generated KIR+NK cells effectively target leukemia cell lines

Acute lymphoid (ALL) and myeloid leukemia (AML) are known to be invasive and highly lethal hematological malignancies. Because current treatments are insufficient and have a variety of side effects, researchers are looking for new and more effective therapeutic methods. Interestingly, ongoing efforts to find the best approach to optimize NK cell anti-leukemia potential shed light on the successful treatment of cancer. Mature KIR+NK cells ability to remove HLA Class-I deficient cells has been exploited in cancer immunotherapy. Here, we generated KIR+NK cells from cord blood stem cells using IL-2 and IL-15 cytokines. Our finding underlined the importance of KIR expression in the cytotoxic function of NK cells. Taken together, this study presented an effective in vitro method for the expansion and differentiation of KIR+NK cells using cytokines without any feeder cells. Furthermore, the presented culture condition could be useful for the generation of mature and pure NK cells from limited numbers of CD34+ cord blood cells and might be used as a novel method to improve the current state of cancer therapy.

A study to assess the knowledge and attitude regarding uses of umbilical cord blood storage among antenatal mothers in selected rural areas at Tumkur district with a view to develop an information booklet

A study to assess the knowledge and attitude regarding uses of umbilical cord blood storage among antenatal mothers in selected rural areas at Tumkur district with a view to develop an information booklet

Combinatorial gene targeting in primary human hematopoietic stem and progenitor cells

The CRISPR/Cas9 system offers enormous versatility for functional genomics but many applications have proven to be challenging in primary human cells compared to cell lines or mouse cells. Here, to establish a paradigm for multiplexed gene editing in primary human cord blood-derived hematopoietic stem and progenitor cells (HSPCs), we used co-delivery of lentiviral sgRNA vectors expressing either Enhanced Green Fluorescent Protein (EGFP) or Kusabira Orange (KuO), together with Cas9 mRNA, to simultaneously edit two genetic loci. The fluorescent markers allow for tracking of either single- or double-edited cells, and we could achieve robust double knockout of the cell surface molecules CD45 and CD44 with an efficiency of ~ 70%. As a functional proof of concept, we demonstrate that this system can be used to model gene dependencies for cell survival, by simultaneously targeting the cohesin genes STAG1 and STAG2. Moreover, we show combinatorial effects with potential synergy for HSPC expansion by targeting the Aryl Hydrocarbon Receptor (AHR) in conjunction with members of the CoREST complex. Taken together, our traceable multiplexed CRISPR/Cas9 system enables studies of genetic dependencies and cooperation in primary HSPCs, and has important implications for modelling polygenic diseases, as well as investigation of the underlying mechanisms of gene interactions.

Criteria for Accepting or Rejecting Cord Blood Units

Rare hematopoietic stem cell populations are responsible for the transplantation engraftment process. Umbilical cord blood (UCB) is usually processed to the total nucleated cell (TNC), but not to the mononuclear cell (MNC) fraction. TNC counts are used to determine UCB unit storage, release for transplantation and correlation with time to engraftment [1]. It has been mentioned in the literature that there are several factors that affect harvesting of cord blood stem cells [2]. It was set to analyze the relationship between fetal weight and other factors that led to rejection of CBUs brought to DCRC from public and private donations. In this retrospective study, 375 donations of CBUs from January 2018 to October 2018 at DCRC were recorded. From literature, factors that are usually involved in rejection of CBUs were recorded and analyzed to mark the leading cause of rejection criteria. In this study, it showed that results did not find infants’ birth weight or mother’s age to have any value in rejection. Other factors’ results will be discussed.

Validation of a rapid potency assay for cord blood stem cells using phospho flow cytometry: The IL-3-pSTAT5 assay.

Public cord blood banks (CBBs) are required to measure cord blood units (CBUs) potency before their release, allowing for the identification of units that may be unsuitable for haematopoietic transplantation. We have developed a rapid flow cytometry assay based on the measurement of STAT-5 phosphorylation of CD34+ stem cells in response to IL-3 stimulation. To adapt the assay from a research setting to its implementation within our CBB regulated operations, we proceded with a full method validation and a correlation comparison of the IL-3-pSTAT5 assay results with the colony-forming unit assay (CFU) results. A total of 60 CBUs cryopreserved in vials were analysed by flow cytometry to determine the sensitivity, specificity, intra-assay precision, robustness, reproducibility, and inter-laboratory agreement of the assay. The CFU assay was also done on the same samples for comparison purposes. The assay threshold was established at 50% CD34+CD45+pSTAT5+, which provides a 100% sensitivity and a 98.3% specificity. An average intra-assay CV of 7.3% was determined. All results met our qualitative results acceptance criteria regarding the inter-user and inter-laboratory agreements, IL-3 stimulation time, post-thaw incubation delay and staining time. The IL-3-pSTAT5 assay results correlated well with the total CFU determined using the CFU assay (r2 = 0.82, n= 56). This study shows that our rapid flow cytometry assay can be successfully validated and that the potency data obtained display good sensitivity, specificity and robustness. These results demonstrate the feasibility of implementing this assay within CBB operations, as a validated potency assay.

Relieving DYRK1A repression of MKL1 confers an adult-like phenotype to human infantile megakaryocytes.

Infantile (fetal and neonatal) megakaryocytes (Mks) have a distinct phenotype consisting of hyperproliferation, limited morphogenesis, and low platelet production capacity. These properties contribute to clinical problems that include thrombocytopenia in neonates, delayed platelet engraftment in recipients of cord blood stem cell transplants, and inefficient ex vivo platelet production from pluripotent stem cell–derived Mks. The infantile phenotype results from deficiency of the actin-regulated coactivator, MKL1, which programs cytoskeletal changes driving morphogenesis. As a strategy to complement this molecular defect, we screened pathways with the potential to affect MKL1 function and found that DYRK1A inhibition dramatically enhanced Mk morphogenesis in vitro and in vivo. Dyrk1 inhibitors rescued enlargement, polyploidization, and thrombopoiesis in human neonatal Mks. Mks derived from induced pluripotent stem cells responded in a similar manner. Progenitors undergoing Dyrk1 inhibition demonstrated filamentous actin assembly, MKL1 nuclear translocation, and modulation of MKL1 target genes. Loss-of-function studies confirmed MKL1 involvement in this morphogenetic pathway. Expression of Ablim2, a stabilizer of filamentous actin, increased with Dyrk1 inhibition, and Ablim2 knockdown abrogated the actin, MKL1, and morphogenetic responses to Dyrk1 inhibition. These results delineate a pharmacologically tractable morphogenetic pathway whose manipulation may alleviate clinical problems associated with the limited thrombopoietic capacity of infantile Mks.

Analysis of risk factors and prognosis of cytomegalovirus infection post umbilical cord blood stem cell transplantation in children with primary immunodeficiency diseases

Objective: To investigate the risk factors and outcomes of cytomegalovirus (CMV) infection post umbilical cord blood stem cell transplantation (UCBT) in children with primary immunodeficiency diseases (PID). Methods: Clinical data of 143 PID children who received UCBT in the Children's Hospital of Fudan University from January 2015 to June 2020 were collected retrospectively. CMV-DNA in the plasma was surveilled once or twice a week within 100 days post-UCBT. According to the CMV-DNA test results, children were divided into the CMV-infected group and the CMV-uninfected group. The incidence and risk factors of CMV infection were analyzed. At 1-month post-UCBT, the absolute lymphocyte count, ratio of lymphocyte subsets and immunoglobulin levels were compared between those whose CMV infection developed 1-month later post-UCBT and those not. Mann-Whitney U test and chi-squared test were used for comparision between groups. Kaplan-Meier survival analysis was used to analyze the impact of CMV infection on survival. Results: Among 143 patients, there were 113 males and 30 females, with a age of 14 (8, 27) months at UCBT. Chronic granulomatosis disease (n=49), very-early-onset inflammatory bowel disease (n=43) and severe combined immunodefiency (n=29) were the three main kinds of PID. The rate of CMV infection was 21.7% (31/143), and the time of infection occurring was 44 (31, 49) days post-UCBT. The incidence of recurrent CMV infection was 4.2% (6/143) and refractory CMV infection was 4.9% (7/143).There was no significant difference in the first time CMV-DNA copy and peak CMV-DNA copy during treatment between the recurrent CMV infection group and the non-recurrent CMV infection group (32.8 (18.3, 63.1)×106 vs. 22.5 (13.2, 31.9)×106 copies/L, Z=-0.95, P=0.340;35.2 (20.2, 54.6)×106 vs. 28.4 (24.1, 53.5)×106copies/L, Z=-0.10, P=0.920), so were those between the refractory CMV infection group and non-refractory CMV infection group (21.8 (13.1, 32.2)×106 vs. 25.9 (14.2, 12.2)×106copies/L, Z=-1.04, P=0.299; 47.7 (27.9, 77.6)×106 vs. 27.7 (19.7,51.8)×106copies/L, Z=-1.49, P=0.137). The CMV-infected group accepted more reduced-intensity conditioning (RIC) regimen than the CMV-uninfected group (45.2% (14/31) vs. 25.0% (28/112), χ2=4.76, P<0.05). The rate of CMV-seropositive recipients and Ⅱ-Ⅳ acute graft versus host diseases (aGVHD) are significantly higher in the CMV-infected group than the CMV-uninfected group (100% (31/31) vs. 78.6% (88/112), 64.5% (20/31) vs. 26.8% (30/112), χ2=7.98,15.20, both P<0.05). The follow-up time was 31.6 (13.2, 45.9) months, CMV infection had no effect on overall survival (OS) rate (χ2=0.02, P=0.843). There was significant difference in the survival rate among three groups of refractory CMV infection, non-refractory CMV infection and the CMV-uninfected (4/7 vs.95.8% (23/24) vs. 86.6% (97/112), χ2=5.91, P=0.037), while there was no significant difference in the survival rate among three groups of recurrent CMV infection, non-recurrent CMV infection and the CMV-uninfected (5/6 vs. 88.0% (22/25) vs. 86.6% (97/112), χ2=0.43, P=0.896). Children who developed CMV infection after 30 days post-UCBT had lower absolute count and rate of CD4+ T cells and immunoglobulin G (IgG) level than those in the CMV-uninfected group (124.1 (81.5, 167.6) ×106 vs. 175.5 (108.3, 257.2) ×106/L, 0.240 (0.164, 0.404) vs. 0.376 (0.222, 0.469), 9.3 (6.2, 14.7) vs. 13.6 (10.7, 16.4) g/L, Z=-2.48, -2.12,-2.47, all P<0.05), but have higher rate of CD8+T cells than those in CMV-uninfected group (0.418 (0.281, 0.624) vs. 0.249 (0.154, 0.434), Z=-2.56, P=0.010). Conclusions: RIC regimen, grade Ⅱ-Ⅳ aGVHD and CMV-seropositive recipients are the main risk factors associated with CMV infection in PID patients post-UCBT. Survival rate of children with refractory CMV infection after UCBT is reduced. Immune reconstitution in children after UCBT should be regularly monitored, and frequency of CMV-DNA monitoring should be increased for children with delayed immune reconstitution.

Application of Stem Cell in Human Erectile Dysfunction - A Systematic Review.

IntroductionErectile dysfunction is a health problem that arises from various conditions and causes an impaired quality of life with a significant health burden. Regenerative and stem cell therapies are some of the potential treatments for erectile dysfunction. This study aimed to review the available information in the literature regarding the use of stem cells in the treatment of erectile dysfunction.MethodsThis study is a systematic review conducted based on the PubMed, Google Scholar, Cochrane, and DOAJ databases. Literature searching was conducted in English and included articles from 2000 to 2020.ResultsThe result was a total of 318 articles. Following the elimination process, 9 articles remained in the final analysis. The analyzed studies included 164 patients with erectile dysfunction with various medical conditions. Several stem cell types have been used for treating erectile dysfunction, including mesenchymal stem cell, placental matrix-derived stem cell, mesenchymal stem cell-derived exosome, adipose-derived stem cell, bone marrow-derived mononuclear stem cell, and umbilical cord blood stem cell. Generally, stem cell therapy showed a good efficacy and safety profile, although not enough studies on the protocol, dosage, and mechanism of action.ConclusionStem cell therapy has a good therapeutic potential in erectile dysfunction, the available data from the literature could be the base of usage of stem cells in the treatment of erectile dysfunction although need more research for broader usage.

Inflammatory gut as a pathologic and therapeutic target in Parkinson’s disease

Parkinson’s disease (PD) remains a significant unmet clinical need. Gut dysbiosis stands as a PD pathologic source and therapeutic target. Here, we assessed the role of the gut-brain axis in PD pathology and treatment. Adult transgenic (Tg) α-synuclein-overexpressing mice served as subjects and were randomly assigned to either transplantation of vehicle or human umbilical cord blood-derived stem cells and plasma. Behavioral and immunohistochemical assays evaluated the functional outcomes following transplantation. Tg mice displayed typical motor and gut motility deficits, elevated α-synuclein levels, and dopaminergic depletion, accompanied by gut dysbiosis characterized by upregulation of microbiota and cytokines associated with inflammation in the gut and the brain. In contrast, transplanted Tg mice displayed amelioration of motor deficits, improved sparing of nigral dopaminergic neurons, and downregulation of α-synuclein and inflammatory-relevant microbiota and cytokines in both gut and brain. Parallel in vitro studies revealed that cultured dopaminergic SH-SY5Y cells exposed to homogenates of Tg mouse-derived dysbiotic gut exhibited significantly reduced cell viability and elevated inflammatory signals compared to wild-type mouse-derived gut homogenates. Moreover, treatment with human umbilical cord blood-derived stem cells and plasma improved cell viability and decreased inflammation in dysbiotic gut-exposed SH-SY5Y cells. Intravenous transplantation of human umbilical cord blood-derived stem/progenitor cells and plasma reduced inflammatory microbiota and cytokine, and dampened α-synuclein overload in the gut and the brain of adult α-synuclein-overexpressing Tg mice. Our findings advance the gut-brain axis as a key pathological origin, as well as a robust therapeutic target for PD.

Identification and Re-consent of Existing Cord Blood Donors for Creation of Induced Pluripotent Stem Cell Lines for Potential Clinical Applications.

We aim to create a bank of clinical grade cord blood-derived induced pluripotent stem cell lines in order to facilitate clinical research leading to the development of new cellular therapies. Here we present a clear pathway toward the creation of such a resource, within a strong quality framework, and with the appropriate regulatory, government and ethics approvals, along with a dynamic follow-up and re-consent process of cord blood donors from the public BMDI Cord Blood Bank. Interrogation of the cord blood bank inventory and next generation sequencing was used to identify and confirm 18 donors with suitable HLA homozygous haplotypes. Regulatory challenges that may affect global acceptance of the cell lines, along with the quality standards required to operate as part of a global network, are being met by working in collaboration with bodies such as the International Stem Cell Banking Initiative (ISCBI) and the Global Alliance for iPSC Therapies (GAiT). Ethics approval was granted by an Institutional Human Research Ethics Committee, and government approval has been obtained to use banked cord blood for this purpose. New issues of whole-genome sequencing and the relevant donor safeguards and protections were considered with input from clinical genetics services, including the rights and information flow to donors, and commercialization aspects. The success of these processes has confirmed feasibility and utility of using banked cord blood to produce clinical-grade iPSC lines for potential cellular therapies.

Abstract 33 A Diverse and Sustainable State-Run Public Cord Blood Program

IntroductionThe Arizona Public Cord Blood Program was created in 2011 by the Arizona Biomedical Research Centre, a subsection of the Arizona Department of Health Services, to advance the collection and increase the number of racially and ethnically diverse cord blood units available for transplantation, as well as to promote awareness of the benefits of cord blood stem cells through our educational partner, Save the Cord Foundation (STCF).ObjectiveThe main objective of this study was to create a sustainable program for women of all racial and ethnic backgrounds to have the opportunity to donate cord blood, with the primary goal of transplantation and the secondary goal of providing non-transplantable cord blood units for research. A second objective was to educate the residents of the state of Arizona about cord blood stem cells and the need for their preservation.MethodsA portion of state lottery funds support the program monetarily. Those funds are provided to four partner collection hospitals employing "cord blood consenters," whose responsibility it is to consent patients, assist delivery providers with collections, and package and ship cord blood units to our partner cord blood bank at MD Anderson Cancer Center. There are also two clinical coordinators who educate and train hospital staff on quality collection practices, with special emphasis on the importance of high-volume, sterile collections. STCF provides education across the state to expectant parents, health care providers, schools, and the public about the need for cord blood stem cell donation for transplant and research.ResultsSince 2011, the Arizona Public Cord Blood Program has banked several hundreds of racially and ethnically diverse cord blood units with the National Marrow Donor Program (Figure 1) and has had 80 life-saving cord blood units matched with patients in need around the globe. This innovative program has expanded cord blood awareness and promoted the preservation of cord blood, and it also has resulted in the creation of an economic engine for the state of Arizona that is an attractant for STEM-based businesses and careers.Figure 1.Pie chart showing percentage of collections by race/ethnicity. Abbreviation: AI/AN, American Indian/Alaskan Native.DiscussionA decade later, the Arizona Public Cord Blood Program has proven to be a sustainable model for collecting and providing suitable cord blood units for transplant to diverse patient populations.

Abstract 29 Unprecedented Experience of Cord Blood Banking in Ukraine During Russian Invasion in 2022

IntroductionOn February 24, 2022, Russia started the unprovoked and unjustified attack on the entire territory of Ukraine. Besides tremendous human losses, the war caused the biggest refugee crisis in Europe since World War II. Approximately 7 million people fled the country; women, some pregnant, and children made 90% of them. Cord blood (CB) banking appeared to be an industry especially sensitive to these unprecedented conditions.ObjectiveThe objective of this study was to analyze the experience of CB banking in Ukraine during the Russia-Ukraine war in 2022.MethodsThematic analysis was performed.ResultsDespite the scarce number of hematopoietic stem cell (SC) transplants for blood malignancies performed there, Ukraine has been a country with a developed regenerative medicine program, with dozens of clinical trials applying perinatal tissues SC. Access to SC therapy was one of the main purposes of medical tourists in Ukraine. The Ukraine’s first CB bank launched in 2003 at the Institute of Cell Therapy LLC and was the main supplier of cell and tissue products to state medical institutions in the country. Thus, the consequences of war, such as disrupted logistics (24,000 km of roads and 300 bridges were destroyed during the first 100 days), ruination of hospitals including maternity clinics (almost 700 health facilities were damaged as of June 9, 2022), lack of human staff (men were mobilized for army and women fled the country), and lack of petroleum, made the successful implementation of contracts on CB storage and SC therapy for both Ukrainian and foreign patients sometimes impossible. The lack of liquid nitrogen experienced during the beginning of the war endangered at least 40,000 CB samples stored at Ukrainian CB banks, as well as biological material at fertility clinics.DiscussionThe unprecedented experience of CB bank operation during the war showed that, for safety reasons, CB banking facilities should be located distantly from strategic objects (eg, railways, airports, and military bases), and should have significant reserve storages of liquid nitrogen, generation reserves, bomb shelter. However, having dedicated personnel was the most important factor allowing for the enterprise functioning in force-majeure events.

Transplantation of human cord blood-derived multipotent stem cells (CB-SCs) enhances the recovery of Parkinson in rats

Earlier published research showed that cord blood-derived multipotent stem cells (CB-SCs) exhibited the intrinsic expression of specific transcription factors (e.g., En1, Nurr1 and Wnt1) and seems to be induced to form dopamine neurons in vitro. In this research, we further investigated the therapeutic potential of CB-SCs in 6-hydroxydopamine lesioned Parkinson's disease (PD) rats. The results of PCR analysis showed that CB-SCs could express transcription factors associated with pluripotentiality and dopaminergic differentiation (e.g., Klf4, c-Myc, Nanog, Sox2, Ngn2, and Nurr1). After being transplanted into the striatum and substantia nigra of PD rats, most of CB-SCs (>90%) developed a fate commitment to dopaminergic differentiation, expressed as the expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT). The improvement effect of cell transplantation on dyskinesia in PD rats was better than that in sham control group. Moreover, higher levels of TH protein in brain homogenates further demonstrated that there were more surviving dopamine neurons in the brain of transplanted PD rats. Study concluds, CB SCS transplantation could promote the regeneration of dopamine neurons and behavioral recovery of PD rats.

EGR1 Haploinsufficiency Confers a Fitness Advantage to Hematopoietic Stem Cells Following Chemotherapy

Therapy-related myeloid neoplasms (t-MNs) share many clinical and molecular characteristics with AML de novo in the elderly. One common factor is that they arise in the setting of chronic inflammation, likely because of advanced age or chemotherapy-induced senescence. Here, we examined the effect of haploinsufficient loss of the del(5q) tumor suppressor gene, EGR1, commonly deleted in high-risk MNs. In mice, under the exogenous stress of either serial transplant or successive doses of the alkylating agent N-ethyl-nitrosourea (ENU), Egr1-haploinsufficient hematopoietic stem cells (HSCs) exhibit a clonal advantage. Complete loss of EGR1 function is incompatible with transformation; mutations of EGR1 are rare and are not observed in the remaining allele in del(5q) patients, and complete knockout of Egr1 in mice leads to HSC exhaustion. Using chromatin immunoprecipitation sequencing (ChIP-seq), we identified EGR1 binding sites in human CD34+ cord blood-derived stem and progenitor cells (HSPCs) and found that EGR1 binds genes critical for stem cell differentiation, inflammatory signaling, and the DNA damage response. Notably, in the chromosome 5 sequences frequently deleted in patients, there is a significant enrichment of innate and inflammatory genes, which may confer a fitness advantage in an inflammatory environment. Short hairpin RNA (shRNA)-mediated silencing of EGR1 biases HSPCs toward a self-renewal transcriptional signature. In the absence of EGR1, HSPCs are characterized by upregulated MYC-driven proliferative signals, downregulated CDKN1A (p21), disrupted DNA damage response, and downregulated inflammation-adaptations anticipated to confer a relative fitness advantage for stem cells especially in an environment of chronic inflammation.

A Study to assess the level of knowledge regarding Umbilical cord blood stem cell banking among antenatal mothers in selected hospital, Dehradun

Introduction: Blood left in the umbilical cord and placenta after birth is known as umbilical cord blood stem cell. Umbilical cord blood is taken from the umbilical vein after the birth when cord is cut and clamped. Umbilical cord blood is stored in both private and public cord banks. Umbilical cord blood can be stored for up to 25-75 years. It is collected through a non-invasive procedure after the birth of the child. This procedure is safe and doesn’t harm the mother and her baby. Objectives /Aims: To assess the level of knowledge among antenatal mothers regarding umbilical cord blood stem cell banking and to find the significant association of knowledge and their demographic variables. Method: The research approach for present study is quantitative approach aimed to assess the knowledge regarding the umbilical cord blood stem cell banking among antenatal mothers. The research design used for this study is descriptive design. The study was conducted in selected hospital at Dehradun. The sample size for this study are 90 antenatal mothers. Convenient sampling technique are used for this study. Self- structured questionnaire method are used for data collection. Result: The present study finding revealed that the majority of the antenatal mothers (64.4%) had poor knowledge, (35.5%) had average knowledge and remaining (0%) had good knowledge. The study result revealed that there was statistically significant association between the level of knowledge with their demographic variables. Conclusion: The purpose of the present study was to find out the knowledge regarding the umbilical cord blood stem cell banking among antenatal mothers in selected hospital of Dehradun. Majority of the antenatal mothers had less knowledge regarding umbilical cord blood stem cell banking and there is need to improve it with the help of health education programme.

Mild or Moderate COVID-19 during Pregnancy Does Not Affect the Content of CD34+ Hematopoietic Stem Cells in Umbilical Cord Blood of Newborns

The study included umbilical cord blood samples (n=64) intended for cryogenic storage of hematopoietic stem cells and obtained from patients with a history of mild and moderate forms of COVID-19 during pregnancy. The control group was composed of samples (n=746) obtained from healthy women in labor. A comparative analysis of the volume of cord blood collected, the total leukocyte count, the relative and absolute content of cells with the CD34+/CD45+ phenotype revealed no significant differences between the groups.

A Review on Recent Advances of Cerebral Palsy

This narrative review summarizes the latest advances in cerebral palsy and identifies where more research is required. Several studies on cerebral palsy were analyzed to generate a general idea of the prevalence of, risk factors associated with, and classification of cerebral palsy (CP). Different classification systems used for the classification of CP on a functional basis were also analyzed. Diagnosis systems used along with the prevention techniques were discussed. State-of-the-art treatment strategies for CP were also analyzed. Statistical distribution was performed based on the selected studies. Prevalence was found to be 2-3/1000 lives; the factors that can be correlated are gestational age and birth weight. The risk factors identified were preconception, prenatal, perinatal, and postnatal categories. According to the evidence, CP is classified into spastic (80%), dyskinetic (15%), and ataxic (5%) forms. Diagnosis approaches were based on clinical investigation and neurological examinations that include magnetic resonance imaging (MRI), biomarkers, and cranial ultrasound. The treatment procedures found were medical and surgical interventions, physiotherapy, occupational therapy, umbilical milking, nanomedicine, and stem cell therapy. Technological advancements in CP were also discussed. CP is the most common neuromotor disability with a prevalence of 2-3/1000 lives. The highest contributing risk factor is prematurity and being underweight. Several preventions and diagnostic techniques like MRI and ultrasound were being used. Treatment like cord blood treatment nanomedicine and stem cell therapy needs to be investigated further in the future to apply in clinical practice. Future studies are indicated in the context of technological advancements among cerebral palsy children.