Source Of Cells For Transplantation Research Articles

Distribution of Amniotic Epithelial Cells After Intraportal Infusion in a Rat Model

Human amniotic epithelial cells (hAECs) are increasingly gaining attention as novel sources for cell transplantation. In clinical practice, intraportal infusion is considered one of the leading approaches for transplantation; however, this has not yet been validated for in vivo transplantation of hAECs. Thus, this study aims to investigate the distribution of hAECs post intraportal infusion and compare this distribution with other cell administration routes. Wistar/ST rats were divided into 4 groups (n=3 for each) based on cell administration route: group 1, intraportal; group 2, the spleen; group 3, tail veins; and group 4, penile veins. Subsequently, hAECs (1×107) stained with XenoLight DiR were infused into each recipient. Cell distribution was evaluated using an in vivo imaging system. DiR signals were detected in the rat livers of groups 1 and 2 with those in group 2 being much weaker than those in group 1. Necrosis of small intestine was observed in 2 cases in group 2. DiR signals were detected in the lungs in groups 3 and 4 because of systemic circulation; however, all the animals died within 20 minutes of infusions. Intraportal infusion is potentially applicable for safe and efficient transplantation of hAECs into the liver, whereas hAECs administration via the spleen carries a risk of thrombosis in a narrow portal vein system. Our results also indicate that hAECs administration via the systemic circulation could cause pulmonary embolism in clinical settings.

Generating pancreatic islets organoids: Langerhanoids

The extension of islet transplantation to a wider number of Type 1 diabetic patients is compromised by the scarcity of donors, the reduced ex vivo survival of pancreatic islets and the use of immunosuppressive treatments. Islets of Langerhans isolated from brain-dead donors are currently the only cell source for transplantation. Thus, it is crucial to find an alternative and an abundant source of functional insulin secreting cells not only for clinical use but also for the development of research dedicated to the screening of drugs and to the development of new therapeutic targets. Several groups around the world, including ours, develop 3D culture models as Langerhanoids that closely mimick human pancreatic islets physiology. In this review, we describe recent advances to mimic the pancreatic niche (extracellular matrix, vascularization, microfluidics) allowing better functionality of Langerhanoids.

Generation of Induced Nephron Progenitor-like Cells from Human Urine-Derived Cells.

Background: Regenerative medicine strategies employing nephron progenitor cells (NPCs) are a viable approach that is worthy of substantial consideration as a promising cell source for kidney diseases. However, the generation of induced nephron progenitor-like cells (iNPCs) from human somatic cells remains a major challenge. Here, we describe a novel method for generating NPCs from human urine-derived cells (UCs) that can undergo long-term expansion in a serum-free condition. Results: Here, we generated iNPCs from human urine-derived cells by forced expression of the transcription factors OCT4, SOX2, KLF4, c-MYC, and SLUG, followed by exposure to a cocktail of defined small molecules. These iNPCs resembled human embryonic stem cell-derived NPCs in terms of their morphology, biological characteristics, differentiation potential, and global gene expression and underwent a long-term expansion in serum-free conditions. Conclusion: This study demonstrates that human iNPCs can be readily generated and expanded, which will facilitate their broad applicability in a rapid, efficient, and patient-specific manner, particularly holding the potential as a transplantable cell source for patients with kidney disease.

Recent advances in stem cell therapy for Parkinson’s Disease: A Review

Parkinson's disease (PD) is a degenerative neurological condition characterized by tremor, bradykinesia, and stiffness as cardinal motor characteristics. It's linked to a long-term loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc), resulting in a severe DA shortage in the striatum, which is necessary for motor function. There is presently no cure for Parkinson's disease, and the majority of treatments aimed at reversing dopamine depletion and alleviating symptoms. The transplantation of stem cells or stem cell derived progenitors has highlighted the potential of employing cell-based therapy to replace lost cells in the sick brain, based on promising findings from early experiments. Embryonic stem cells (ESCs) are highly expandable and pluripotent cells that can differentiate into any cell type in the human body, including nervous system tissues, suggesting that they could provide a long-term treatment for Parkinson's disease and other neurological illnesses. However, because of the potential for safety and ethical difficulties involved with the use of undifferentiated ESCs in people, other sources of transplantable cells must be considered. Another method is to use external manipulation to stimulate endogenous stem cells to heal the brain. Recent advances in stem cell research in Parkinson's disease will be discussed in this review, which will provide an overview of the various sources and strategies such as the use of different stem cell populations for cell replacement and possible modulation of endogenous stem cells, that have the potential to provide effective cell-based therapy in the future.

Unexpected High Frequency of Pathogenic Germline Variants in Older Adults with Primary Myelodysplastic Syndrome

Background: Germline predisposition is increasingly being recognised in myeloid neoplasms (MN) including primary myelodysplastic syndrome. An unequivocal diagnosis of germline predisposition carries actionable considerations for patient management including donor stem cell source for allogeneic transplantation, dose-reduction of conditioning regimes and screening for extra-hematological disease (such as pulmonary abnormalities in patients with telomere biology disorders). In addition, the identification of MDS predisposition syndromes can avoid misdiagnosis (for example, distinguishing idiopathic thrombocytopenic purpura from thrombocytopenia due to RUNX1 germline variant). However, the prevalence of pathogenic germline variants (PGVs) in unselected pMDS patients presenting at older age remains unknown.Aim: This study assesses frequency and type of pathogenic germline variants in MDS patients and compares with age matched healthy controls and patients with other cancers.Method: We analysed 68 known cancer predisposition genes in germline samples of 146 samples from myeloid neoplasms. Study included primary MDS (n=51) and MDS diagnosed in cancer survivors with (n=77) or without prior exposure to cytotoxic therapy (n=18). Using uniform American College of Medical Genetics and Genomics (ACMG) guidelines for annotating germline mutation, we also compared the frequency of pathogenic germline variants in the same genes with patients with single cancer and age-match healthy controls (>70 years).Results: Pathogenic germline variants (PGVs) were identified in 19% (28/146) patients compared to 4% and 3% patients with single cancer and age-matched controls respectively (P<0.0001) (Fig. 1A). Median age at diagnosis was similar between MN patients with or without PGVs [66 years (19-81) vs. 70 years (33-87); P=0.06]. PGVs were most frequent in DDX41 (n=7, 33%) followed by BRCA1 (n=2, 10%), GATA2 (n=2; 10%) and TP53 (n=2; 10%) (Fig.1B). We also identified pathogenic copy number variations (CNV) in 4 patients. The distribution of PGVs was also different, with DDX41 PGVs absent in single cancers and more prevalent in MN than age-matched controls (35% vs. 4%, P<0.001). The frequency of PGV was not significantly different between P-MN and T-MN/ MC-MN (17% versus 10%, P = 0.32 (Fig.1C). The frequency of PGV was 30%, 6%, 19%, 15% and 18% in patients ≤50, 51-59, 60-69, 70-79 and >80 years of age (Fig. 1D).Phenotypic features such as monocytopenia and mycobacterium infections (MonoMAC; SA460) and personal and family history of pulmonary fibrosis (SA918) were present in only two cases with PGVs. Family history of MDS/AML was present in only in four cases with PGVs, in which PGVs were found in typical myeloid malignancy genes (DDX41, GATA2). Importantly, some patients with family history of solid cancers carried PGVs in genes traditionally associated with solid cancers (e.g. MSH6, NF1, TP53 and BRCA1). SA927 had a PGV in MSH6 and multiple first-degree relatives with solid cancers including colon, renal and brain cancers. Moreover, 41% of adults with hematological disorders and a personal and/or family history of interstitial lung disease had PGVs in telomere biology disorder genes. Hence, family history should not be restricted to hematological disorders, but also solid cancers and non-malignant phenotypes (e.g. hepatic and pulmonary fibrosis). The frequency of PGVs was not different in patients with and without family history of cancer (23% vs. 13%, P=0.32).Conclusion: The frequency of PGVs is significantly high in MN compared to age matched healthy control and other cancer patients. Our observation of a high frequency of PGVs in the older MDS population warrants standardization of germline testing at diagnosis to guide optimal management of patients and their families. [Display omitted] DisclosuresHiwase: Novartis: Membership on an entity's Board of Directors or advisory committees; AbbVie: Membership on an entity's Board of Directors or advisory committees.

Role of the Interaction Between Nck1 and PERK in Pancreatic Beta Cell Function and Survival

Preventing pancreatic β cell failure and death, features of diabetes pathogenesis, could help treat diabetes. PERK, a player in the unfolded protein response (UPR), is essential for β cell function and survival. We have shown that PERK, when residue Y561 is phosphorylated, is bound and regulated by the adaptor protein Nck1 (non-catalytic region of tyrosine kinase adaptor protein 1). Silencing Nck1 in β cells enhances basal PERK activity and prevents its hyperactivation under gluco-lipotoxic conditions, improving proinsulin synthesis and β cell survival. Our preliminary results also showed that islet grafts lacking Nck1 perform better compared to WT grafts to reverse streptozotocin-induced diabetes in mice. Thus, we hypothesize that disruption of the specific Nck1/PERK interaction could improve β cell health and generate a phosphopeptide (TAT-pY561) that sequesters Nck1 and prevents its interaction with PERK. As predicted, this peptide increases insulin synthesis and decreases apoptosis in cultured beta cells under gluco-lipotoxic conditions. In vivo, the peptide administered daily for 3 weeks to mice (prior to and during induction of diabetes using HFD + streptozotocin) increased pancreatic insulin content and glucose-stimulated insulin secretion compared to vehicle treated mice. Moreover, in human diabetic islets, in vitro treatment with the peptide tended to increase insulin content. Our data suggest that inhibition of Nck1/PERK interaction, in mouse and human islets, improves β cell survival and function. Targeting this interaction might prove useful to enhance beta cell health in diabetes and/or when producing sources of beta cells for transplantation.

Machine learning–based scoring models to predict hematopoietic stem cell mobilization in allogeneic donors

AbstractMobilized peripheral blood has become the primary source of hematopoietic stem cells for both autologous and allogeneic stem cell transplantation. Granulocyte colony-stimulating factor (G-CSF) is currently the standard agent used in the allogeneic setting. Despite the high mobilization efficacy in most donors, G-CSF requires 4–5 days of daily administration, and a small percentage of the donors fail to mobilize an optimal number of stem cells necessary for a safe allogeneic stem cell transplant. In this study, we retrospectively reviewed 1361 related allogeneic donors who underwent stem cell mobilization at Washington University. We compared the standard mobilization agent G-CSF with five alternative mobilization regimens, including GM-CSF, G-CSF+GM-CSF, GM-CSF + Plerixafor, Plerixafor and BL-8040. Cytokine-based mobilization strategies (G-CSF or in combination with GM-CSF) induce higher CD34 cell yield after 4–5 consecutive days of treatment, while CXCR4 antagonists (plerixafor and BL-8040) induce significantly less but rapid mobilization on the same day. Next, using a large dataset containing the demographic and baseline laboratory data from G-CSF–mobilized donors, we established machine learning (ML)–based scoring models that can be used to predict patients who may have less than optimal stem cell yields after a single leukapheresis session. To our knowledge, this is the first prediction model at the early donor screening stage, which may help identify allogeneic stem cell donors who may benefit from alternative approaches to enhance stem cell yields, thus ensuring safe and effective stem cell transplantation.

Assessment of HLA-B genetic variation with an HLA-B leader tool and implications in clinical transplantation

AbstractSequence variation in the HLA-B gene is critically linked to differential immune responses. A dimorphism at -21 of HLA-B exon 1 gives rise to leader peptides that are markers for risk of acute graft-versus-host disease, relapse, and mortality after unrelated donor and cord blood transplantation. To optimize the selection of stem cell transplant sources based on the HLA-B leader, an HLA-BLeader Assessment Tool (BLEAT) was developed to automate the assignment of leader genotypes, define HLA-B leader match statuses, and rank order candidate stem cell sources according to clinical risk. The base cohort consisted of 9 417 614 registered donors from the Be The Match Registry with HLA-B typing. Among these donors, the performance of BLEAT was assessed in 1 098 358 donors with sequence data for HLA-B exon 1 (2 196 716 haplotypes). The accuracy of leader assignment was then assessed in a second cohort of 1259 patients and their unrelated transplant donors. We furthermore established the frequencies of HLA-B leader genotype (MM, MT, TT) representations in broad racial categories in the 9.42 million donors. BLEAT has direct applications for the selection of optimal stem cell sources for transplantation and broad utility in basic and clinical research in pharmacogenomics, vaccine development, and cancer and infectious disease studies of human populations.

Stem Cells for Aging-Related Disorders.

This review captures recent advances in biological and translational research on stem cells. In particular, we discuss new discoveries and concepts regarding stem cell treatment of aging-related disorders. A myriad of stem cell sources exists, from hematopoietic to mesenchymal and neural cell lineages. We examine current applications of exogenous adult bone marrow-derived stem cells as an effective and safe transplantable cell source, as well as the use of electrical stimulation to promote endogenous neurogenesis for Parkinson's disease. We also explore the potential of transplanting exogenous umbilical cord blood cells and mobilizing host resident stem cells in vascular dementia and aging. In addition, we assess the ability of small molecules to recruit resident stem cells in Alzheimer's disease. Finally, we evaluate mechanisms of action recently implicated in stem cell therapy, such as the role of long non-coding RNAs, G-protein coupled receptor 5, and NeuroD1. Our goal is to provide a synopsis of recent milestones regarding the application of stem cells in aging.

TIPE2-modified human amnion-derived mesenchymal stem cells promote the efficacy of allogeneic heart transplantation through inducing immune tolerance.

BackgroundImmune rejection of heart transplantation has been regarded as the biggest challenge encountered by a patient suffering from end-stage heart disease. The transplantation of human amnion-derived mesenchymal stem cells (hAD-MSCs) has exhibited promising application prospects in organ transplantation. However, its persistent unsatisfactory tolerance has limited the widespread application of this technology. We aim to investigate the role of tumor necrosis factor-α-induced protein-8 like-2 (TIPE2)-mediated hAD-MSCs in immune tolerance in heart transplantation and its molecular regulatory mechanisms.MethodsThis project detected the effect of TIPE2 on immune tolerance by constructing an allogeneic heart transplantation mouse model through which TIPE2-overexpressed hAD-MSCs were injected into recipients. The fluorescence distribution of TIPE2-hAD-MSCs in mice was observed by a small animal in vivo imaging system. Pathological changes of the transplanted heart were detected by hematoxylin and eosin (HE) staining. Flow cytometry was performed to detect the content of cardiac lymphocytes. The expression of immune-induced related factors was measured by quantitative real-time PCR (qRT-PCR) and western blot assays.ResultsTIPE2-hAD-MSCs protected myocardial tissue structures, reduced the spleen and thymus indexes in recipient mice, minimized the content of cardiac lymphocytes, reduced expressions of ERK, p38, and IFN-γ, and elevated expressions of both IL-10 and TGF-β, markedly improving the survival time and survival rates of recipient mice.ConclusionsTIPE2-hAD-MSCs induce immune tolerance and improve the survival rates of allogeneic heart transplantation in mice. This study is expected to offer an ideal source and target of cells for organ transplantation.

Optimal umbilical cord blood collection, processing and cryopreservation methods for sustained public cord blood banking

Background aimsUmbilical cord blood is an established source of stem cells in patients with hematologic malignancies who do not have HLA-compatible matched related or unrelated donors. The success of an umbilical cord blood transplant depends on the dose of total nucleated and CD34+ cells infused. Therefore, collecting, banking and listing high-quality cord blood units with high total nucleated and CD34+ cell dose are essential. MethodsHere the authors describe their cord blood bank's novel collection technique, which involves both in utero and ex utero collection of a single cord blood unit. The authors also evaluated maternal, neonatal and collection parameters that may impact the cell dose. ResultsMaternal gestational age and race, and neonatal weight and sex correlated with the total nucleated cell dose. ConclusionsThe optimized collection of umbilical cord blood is critical for its use as a source of stem cells for transplantation.

Assessment of a Multi-Cytokine Profile By a Novel Biochip-Based Assay Reveals Unique Correlation Patterns with Immune Reconstitution after Cord Blood Transplantation in Adults

Umbilical cord blood transplantation (UCBT) is an alternative stem cell source for allogeneic hematopoietic stem cell transplantation. The frequency of acute graft-versus-host-disease (aGvHD) in UCBT recipients is less than expected in recipients of adult donor allografts but delayed aGvHD can develop. Moreover, UCBT results in delayed and insufficient immune reconstitution leading to infection-related morbidity and mortality. We reasoned that an approach that would allow risk assessment for development of GvHD and/or for impaired immune reconstitution would be highly beneficial in adult UCBT recipients because it would guide therapeutic decisions for the prevention of these complications. Because reconstitution of hematopoietic lineages is delayed in UCB recipients, we focused on cytokines that can be assessed in patients' plasma at any time point before or after UCBT. Increase of proinflammatory cytokines IL-1b and TNF-α after BMT has been associated previously with the pathogenesis of GvHD. We previously determined that levels of IL-7 were inversely correlated with reconstitution of T cell subsets but were also associated with non-relapse mortality, development of GvHD and worse overall survival. In the present study, we employed a novel approach, which allows simultaneous assessment of multiple cytokines in a very small volume of patient's serum or plasma, to assess the prognostic value of this multi-cytokine profile in adult UCBT recipients. 32 patients with a median age of 50 years with hematopoietic malignancies were treated with reduced intensity conditioning (Flu/Mel/rATG) followed by two sequential UCB graft transplantation (dUCBT) and GvHD prophylaxis with tacrolimus and sirolimus. Results are based on 27 evaluable patients. Assessments were done prior to transplantation and at 1, 2, 3, 6, 12 and 24 months after dUCBT. Quantification of cytokines IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IFN-γ, TNF-α and GM-CSF in serum samples was conducted using the LUNARISTM Human 11-Plex Cytokine Biochip384 from AYOXXA Biosystems. Briefly, after diluting as 1:2 using the respective assay diluent, 5µl of samples or assay standards were loaded in duplicates on the AYOXXA Biochip and the assay was performed according to the manufacturers' instructions. Readout was performed using the AYOXXA Reader AR01 (Cat. No. LRS-001, AYOXXA Biosystems) and data were analyzed using the LUNARISTM Analysis Suite Software (Cat. No. LAS-001, AYOXXA Biosystems). Based on their median levels, cytokines could be categorized in four patterns of kinetics in comparison to pre-transplant baseline values: 1) Without significant changes (IL-2); 2) decrease over time (IL-1b, IL-17); 3) increase at 1-month after dUCBT and subsequent decline (IL-4, IL-5, IL-6, IL-8, IL-10, TNF-α, GM-CSF); 4) increase at 2 months after UCBT and subsequent decline (IFN-γ). At baseline, most cytokines correlated with total lymphocytes, CD4+, CD8+ subsets, CD4+CD25+ Treg, CD56+CD16+ NK cells, CD20+ B cells and CD14+ monocytes. At 1-month after dUCBT, the correlations with immune cell types disappeared for most cytokines and only CD14+ monocytes correlated with IFN-γ. At 2-months after dUCBT, most cytokines were positively correlated with T cell subsets. IL-8 displayed a unique pattern of correlation with CD14+ monocytes and B cells, both of which have potent antigen presenting function. Specifically, at 2-months after dUCBT IL-8 inversely correlated with CD14+ cells (r=-0.71, p=0.0007) and this pattern persisted until day 100, when CD20+ cells also showed a significant inverse correlation with IL-8 (r=-0.56, p=0.006). These inverse correlations declined by 6 months after dUCBT. IL-8 levels after BMT were previously associated with infectious complications, which subsequently drive elevation of IL-6 and development of GvHD. Our findings pose the intriguing possibility that multicytokine profile analysis might serve as a biomarker to guide patient management such as work-up for infectious complications that might prevent GVHD development in dUCBT recipients. Analysis of correlations between multicytokine profile and such clinical outcomes is currently ongoing. Our results provide evidence for the feasibility of a novel method for multicytokine assessment in patients' plasma, which might guide patient management, to diminish complications after dUCBT. [Display omitted] DisclosuresAntin:Gentium SpA/Jazz Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees.

Hypoxia Preconditioning Promotes the Proliferation and Migration of Human Urine-Derived Stem Cells in Chronically Injured Liver of Mice by Upregulating CXCR4.

Our previous studies reported that urine-derived stem cells (USCs) possess a strong self-renewal ability and multidirectional differentiation potential and thus are an ideal candidate cell source for hepatocellular transplantation. USC transplantation may repair the pathological changes of chronic liver injury to a certain extent, and hypoxia pretreatment may improve the recovery efficiency of USCs. Therefore, the present study aimed to investigate the possible mechanism of the improved recovery efficiency of hypoxia-pretreated USCs. A chronic liver injury model was established by intraperitoneal injection of carbon tetrachloride into nude mice. USCs were transplanted via caudal vein injection. Hematoxylin and eosin staining and Masson's staining were performed to determine the pathology of the liver. Immunofluorescence and frozen section biopsy were performed to determine differentiation and cell fusion in vivo. Cell coculture was used to detect cell fusion in vitro. The proliferative ability of USCs was evaluated using cell viability and colony formation assays, and the migratory functions of USCs were evaluated using wound healing and transwell assays. The degeneration of hepatocytes and the level of fibrosis in the hypoxia transplantation group were improved compared with the normoxia transplantation group. It was found that exogenous USCs may be differentiated into functional hepatocytes or fused with hepatocytes in vivo. C-X-C motif chemokine (CXC) ligand 12 (CXCL12) expression levels in liver tissue of the chronic liver injury model were upregulated compared with those in the control group. The expression of CXC receptor 4 (CXCR4) in hypoxia-pretreated USCs was also significantly upregulated. The results suggested that USCs fused with different types of liver cells and that hypoxia treatment promoted the fusion rate in vitro by upregulating CXCR4 signaling. Furthermore, hypoxia pretreatment promoted cell proliferation, migration, and cell fusion by inducing CXCR4 signaling, leading to USC-elicited liver tissue recovery following injury in vivo.

Control of Microbial Opsin Expression in Stem Cell Derived Cones for Improved Outcomes in Cell Therapy.

Human-induced pluripotent stem cell (hiPSC) derived organoids have become increasingly used systems allowing 3D-modeling of human organ development, and disease. They are also a reliable source of cells for transplantation in cell therapy and an excellent model to validate gene therapies. To make full use of these systems, a toolkit of genetic modification techniques is necessary to control their activity in line with the downstream application. We have previously described adeno-associated viruse (AAV) vectors for efficient targeting of cells within human retinal organoids. Here, we describe biological restriction and enhanced gene expression in cone cells of such organoids thanks to the use of a 1.7-kb L-opsin promoter. We illustrate the usefulness of implementing such a promoter to enhance the expression of the red-shifted opsin Jaws in fusion with a fluorescent reporter gene, enabling cell sorting to enrich the desired cell population. Increased Jaws expression after transplantation improved light responses promising better therapeutic outcomes in a cell therapy setting. Our results point to the importance of promoter activity in restricting, improving, and controlling the kinetics of transgene expression during the maturation of hiPSC retinal derivatives. Differentiation requires mechanisms to initiate specific transcriptional changes and to reinforce those changes when mature cell states are reached. By employing a cell-type-specific promoter we put transgene expression under the new transcriptional program of mature cells.

Towards a Functional Cure for Diabetes Using Stem Cell-Derived Beta Cells: Are We There Yet?

Diabetes mellitus is a pandemic metabolic disorder that results from either the autoimmune destruction or the dysfunction of insulin-producing pancreatic beta cells. A promising cure is beta cell replacement through the transplantation of islets of Langerhans. However, donor shortage hinders the widespread implementation of this therapy. Human pluripotent stem cells, including embryonic stem cells and induced pluripotent stem cells, represent an attractive alternative beta cell source for transplantation. Although major advances over the past two decades have led to the generation of stem cell-derived beta-like cells that share many features with genuine beta cells, producing fully mature beta cells remains challenging. Here, we review the current status of beta cell differentiation protocols and highlight specific challenges that are associated with producing mature beta cells. We address the challenges and opportunities that are offered by monogenic forms of diabetes. Finally, we discuss the remaining hurdles for clinical application of stem cell-derived beta cells and the status of ongoing clinical trials.

First trimester placental mesenchymal stem cells improve cardiac function of rat after myocardial infarction via enhanced neovascularization

Acute myocardial infarction (AMI) is the most critical heart disease. Mesenchymal stem cells (MSCs) have been widely used as a therapy for AMI for several years. The human placenta has emerged as a valuable source of transplantable cells of mesenchymal origin that can be used for multiple cytotherapeutic purposes. However, the different abilities of first trimester placental chorion mesenchymal stem cells (FCMSCs) and third trimester placental chorion mesenchymal stem cells (TCMSCs) have not yet been explored. In this study, we aimed to compare the effectiveness of FCMSCs and TCMSCs on the treatment of AMI. FCMSCs and TCMSCs were isolated and characterized, and then they were subjected to in vitro endothelial cell (EC) differentiation induction and tube formation to evaluate angiogenic ability. Moreover, the in vivo effects of FCMSCs and TCMSCs on cardiac improvement were also evaluated in a rat MI model. Both FCSMCs and TCMSCs expressed a series of MSCs surface markers. After differentiation induction, FCMSCs-derived EC (FCMSCs-EC) exhibited morphology that was more similar to that of ECs and had higher CD31 and vWF levels than TCMSCs-EC. Furthermore, tube formation could be achieved by FCMSCs-EC that was significantly better than that of TCMSCs-EC. Especially, FCMSCs-EC expressed higher levels of pro-angiogenesis genes, PDGFD, VEGFA, and TNC, and lower levels of anti-angiogenesis genes, SPRY1 and ANGPTL1. In addition, cardiac improvement, indicated by left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVEDs), left ventricular ejection fraction (LVEF) and left ventricular shortening fraction (LVSF), could be observed following treatment with FCMSCs, and it was superior to that of TCMSCs and Bone marrow MSCs (BMSCs). FCMSCs exhibited a superior ability to generate EC differentiation, as evidenced by in vitro morphology, angiogenic potential and in vivo cardiac function improvement; further, increased levels of expression of pro-angiogenesis genes may be the mechanism by which this effect occurred.

Feeder-Free Cell Culture of Labial Oral Mucosal Epithelium for Tissue-Engineering and Regenerative Medicine

Background.The cultured cheek mucosa epithelium (buccal epithelium, BE) is used for autologous transplants generation and tissue engineering. An alternative source of cells for these purposes may be the lip mucosa, covered, like BE, with non-keratinized stratified squamous epithelium, but with some histological distinctions.Aims to characterize the human lip mucosa as a promising source of epithelial cells for autologous transplantation and tissue engineering.Methods.Scrapings of the lip, cheek, and gum mucosa from five healthy volunteers were analyzed by cytofluorimetry to determine the level of desquamation and cytokeratin (CK) 10 and 13 expression. The lip mucosa of two patients was characterized using routine histological staining and fluorescence immunohistochemistry for CK 3, 4, 10, 13, and p63 marker. 35 samples of full-thickness strips of the patients lip mucosa were used to set the explant (n=18) and enzymatic (n=17) techniques for expansion epithelium. Culture systems with 1.05 and 0.06mM Calcium contained 5% fetal bovine serum, 5 g/ml human insulin, 5 g/ml hydrocortisone, 10 ng/ml human epidermal growth factor. Stable cultures were stained for p63, vimentin, zonula occludens-1 (ZO-1), and CK10. Software tools determined levels of their expression.Results.The number of cells in the lip and gum samples was significantly lower than from the cheek. The median number of CK13 positive cells was significantly different for the gum (6.4%) and cheek (64.8%, p=0.0089). Significant differences for CK10 positive cells were not observed. The epithelium of the lip mucosa was 72.13.6 m thick, relatively flat, and without keratinization sites. Samples were positively stained for CK 4 and 13, in the absence of expression of CK 3 and 10. The primary culture of epithelial cells obtained by explant technique was significantly more effective (p=0.001) in comparison with the enzymatic method. Stable cultures had a cobble-stone morphology in both culture systems. The levels of vimentin and p63 expression in both culture systems was not significantly differ. ZO-1 expression was 3.6-fold higher for 1.05-mMCa++medium (p=0.0006).Conclusions.Epithelium cell culture from the lip mucosa can be obtained by culturing explants without a feeder layer. Quality control steps have been developed for cultured cells and biopsy site.

Blood stem cells compared with bone marrow as a source of hematopoietic cells for allogeneic transplantation

AbstractPeripheral blood cells are increasingly used in place of bone marrow as a source of hematopoietic stem cells for allogeneic transplantation. The relative efficacy of these 2 approaches is unknown. This retrospective multivariate analysis compared results of 288 HLA-identical sibling blood stem cell transplantations with results of 536 HLA-identical sibling bone marrow transplantations. No transplants were T-cell depleted. Median follow-up was 12 months, and analyses focused on 1-year outcomes. Recipients of blood stem cell transplants had more rapid recovery of neutrophils to at least 0.5 × 109/L (median time to recovery, 14 days, compared with 19 days for marrow transplants;P < .001) and of platelets to at least 20 × 109/L (median time, 18 days, compared with 25 days for marrow transplants; P < .001). There was no significant difference in the incidence of grades II to IV acute graft versus host disease (GVHD). The incidence of chronic GVHD was significantly higher after blood stem cell transplantation (1-year probability [95% confidence interval], 65% [56%-72%] compared with 53% [47%-59%]; P = .02) Relapse incidence in the 2 transplant groups did not differ significantly. Treatment-related mortality rates were lower and leukemia-free survival rates were higher with blood stem cell transplants in patients with advanced leukemia (acute leukemia in second remission or chronic myelogenous leukemia in accelerated phase) but not in early leukemia (acute leukemia in first remission or chronic myelogenous leukemia in chronic phase). The median time from transplantation to hospital discharge was 23 days after blood stem cell transplantation and 28 days after bone marrow transplantation (P = .003). Further study with longer follow-up is necessary to definitively establish the role of blood stem cells for allogeneic transplantation, especially in patients with good-risk disease.

Analysis of the legal regulation of the use of umbilical cord blood and its components in the Russian Federation and abroad

Umbilical cord blood is a unique source of hematopoietic cells for transplantation in hematological diseases and other socially significant pathologies. The development of regenerative medicine requires clear protocols governing the use of umbilical cord blood and its components (cells, plasma) in clinical practice. In the Russian Federation, today, there is no proper regulatory framework for working with umbilical cord blood and its components, which is a limiting factor in the development of cellular technologies and regenerative medicine in general. The article analyzes the legal framework for regulating the use of umbilical cord blood and its components in the Russian Federation and abroad - the countries of the European Union and America.

Слизистая губы как источник эпителиальных клеток для аутологичной трансплантации при синдроме лимбальной недостаточности роговицы

Слизистая губы как источник эпителиальных клеток для аутологичной трансплантации при синдроме лимбальной недостаточности роговицы