Source Of Stem Cells Research Articles

Stromal Vascular Fraction of Adipose Tissue: Prospects for Use in the Treatment of Osteoarthritis and Articular Cartilage Injuries (Literature Review)

Summary. Osteoarthritis (OA) is a chronic, progressive, degenerative-dystrophic joint disease primarily caused by damage to the articular cartilage. Conservative pharmacological treatment of OA consists of prescribing nonsteroidal anti-inflammatory drugs, chondroprotectors, hyaluronic acid injections, and therapeutic blockades with glucocorticosteroids. The use of orthobiological agents represents an alternative method in OA treatment. Among the variety of orthobiological approaches, adipose tissue serves as a new and attractive source of stem cells for the treatment of OA. The stromal vascular fraction creates an environment where stem cells can interact and stimulate regeneration.

Outcome of hematopoietic stem cell transplantation for SCID and impact of newborn screening on overall survival - A single referral center study.

Hematopoietic stem cell transplantation (HSCT) is a curative treatment for infants with severe combined immunodeficiency (SCID). Different factors determine HSCT success and overall survival (OS). Specifically, prompt diagnosis of SCID, preferably through newborn screening (NBS), is critical. To explore variable factors including the impact of NBS that are associated with HSCT outcomes and OS, in a multi-ethnic SCID cohort. 100 SCID patients diagnosed and treated with HSCT at Sheba Medical Center in Israel between the years 1996 to 2024 were studied. We distinguished three groups: Palestinians (62%), Israeli Jews (16%), and Israeli non-Jews (22%). The OS rate was 68%, increasing to 84% when excluding mortality in the first month post-transplantation. Better outcomes were significantly associated with the use of bone marrow (BM) as a stem cell source (P =0.003), the availability of matched related donors (MRD, P =0.045) and use of conditioning (P =0.0006). Due to delayed diagnosis, Palestinian patients had more infections, more events of early post-HSCT death, and inferior OS rates compared to other patients. SCID cases identified by NBS demonstrated superior OS (93%) compared to cases identified by clinical presentation (P =0.04). Improvement in OS was most significant following the implementation of the NBS program for SCID in Israel (P = 0.03). Our study delineates and reinforces specific factors that influence OS after undergoing HSCT for SCID. Importantly, it raises the value of early diagnosis and treatment of affected infants, highlighting the benefit of NBS for SCID in determining the clinical outcome.

Haploidentical Stem Cell Transplantation With Dual Source of Cells and Post-Transplant Cyclophosphamide.

Dual sources of cells (DSC) with peripheral blood stem cell apheresis (PBSC) and surgical bone marrow (BM) for haploidentical hematopoietic cell transplantation (Hid-HCT) are used in China and some Asian countries. The experience of the Baltimore group for haploidentical transplant with post-transplant cyclophosphamide (PT-Cy) and reduced-intensity-conditioning (RIC) regimen used BM as a source of hematopoietic stem cells. We retrospectively analyzed 64 Hid-HCT with DSC and PT-Cy, RIC (n = 57), or myeloablative-conditioning (MAC) (n = 7), from two public health Brazilian centers, with a median follow-up of 23.3 months (6.7-45.4). The 49 malignant patients were 27/46 (58.7%) beyond the first remission or with no complete response, and three patients did not complete disease status evaluation before transplant. Eight of 62 patients (12.9%) had grade 2 or more Hematopoietic Cell Transplantation-Specific Comorbidity Index (HCT-CI), and two patients had no HCT-CI classified. Cytomegalovirus (CMV) viremia occurred in 26 of 57 (45.6%). The cumulative incidence of 100-day grade III-IV acute GVHD was 12.3% (7/57), with a 95% confidence interval (CI) of 3.8% and 20.8%, and 2-year moderate or severe chronic GVHD was 21.1% (11/52; 95% CI, 10.1%-32.3%). The 2-year relapses were 24.5% for malignant disease (12/49; 95% CI, 12.4%-36.5%). The 2-year overall survival (OS) probability was 54.7% (35/64; 95% CI, 42.5%-66.9%). Benign diseases achieve 2-year OS in 73.3% (11/15; 95% CI, 51%-95.7%) of the patients. The HCT-CI were significant in multivariate analyses for DFS (p = 0.002) and OS in uni- and multivariate analyses (both p < 0.001). The number of CD34+ cells by apheresis collection was significant in multivariate analysis for DFS (p = 0.039). Hid-HCT using PT-Cy, DSC, and RIC is a safe option for benign and malignant diseases.

P-2356. A SFGM-TC real-world study to characterize pediatric patients undergoing allogeneic Hematopoietic Stem Cell Transplantation (HSCT) and identify the cytomegalovirus (CMV) serology status of both recipients and their donors

Abstract Background The aim of this study was to perform a description of pediatric patients’ characteristics undergoing an allogeneic Hematopoietic Stem Cell Transplantation (HSCT) and to identify their cytomegalovirus (CMV) serology status and that of their donors Methods Patients under the age of 18 who received an allogeneic HSCT in a French center reporting data in the ProMISe database of the francophone society of bone marrow transplant and cellular therapy (SFGM-TC) during the period comprised between January 1, 2019, and December 31, 2021 Results 904 allo-HSCT were performed between January 1, 2019, and December 31, 2021, in 22 hematopoietic stem cell transplant centres in France. Among them 57,5% (520) were males (Figure 1). Patients ages were analysed with 14,7% (133) being strictly below 2 years old, 29,8% (269) being between 2 and 6 years old, 25,9% (234) being between 7 and 11 years old and 29,7% (268) being between 12 and 18 years old. Age classes distribution wasn’t different throughout the years analysed, as shown in Figure 2 CMV serostatus of both donor and recipient were available for 96% of patients. Donor negative and Recipient negative (D-/R-) accounted for 32% of the population, Donor positive and Recipient negative (D+/R-) accounted for 16%, Donor Positive and Recipient positive (D+/R+) accounted for 29% and lastly Donor negative and Recipient positive (D-/R+) accounted for 19%. Serostatus was analysed through age classes (Table 1) showing a numerical trend toward a higher CMV seropositivity for recipient being older than 2 years old. R+ patients were 42,1% below 2 years old and 49,3% in the 2 to 6-year-old age group. The percentage remained stable in older patients. The primary diagnoses leading to the graft were mainly primary immune deficiency for patients under 2 years old, accounting for 45.1% (60/133). After 2 years old, the leading cause was acute leukaemia (Table 2). Pediatric patients mainly received bone marrow (71%) as the source of stem cells, followed by cord blood (18%) and peripheral blood (11%).Table 1CMV Serostatus according to age of recipients Conclusion The diagnosis leading to HSCT in the pediatric population differs according to age class. Serostatus also differs according to age class, with older patients approaching the levels of seropositivity seen in adults. Disclosures Xavier Bourge, PharmD, MSD France: Employee Guilhem Tournaire, MD, MSD France: Employee Jean-Hugues Dalle, MD, MSD France: Advisor/Consultant Bénédicte Neven, MD, MSD France: Advisor/Consultant Marie Robin, MD, MSD France: Grant/Research Support Raynier DEVILLIER, MD, MSD France: Grant/Research Support Nicole Raus, n/a, MSD France: Grant/Research Support Stephanie Nguyen, n/a, MSD France: Grant/Research Support

Advances in allogeneic hematopoietic stem cell transplantation for Langerhans cell histiocytosis in children

Langerhans cell histiocytosis (LCH) is a disease caused by clonal expansion of CD1a+/CD207+ cells and is characterized by organ involvement and dysfunction. Treatment of LCH in children is risk-adapted, and multisystem LCH requires systemic therapy. Although systemic treatments such as chemotherapy and BRAF/MEK inhibitors have improved the cure rate of LCH, disease reactivation rates remain 30%, and eventually some patients progress to relapse-refractory LCH. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a promising salvage treatment strategy for children with relapse-refractory LCH. However, many questions such as the efficacy and indications of allo-HSCT, as well as suitable conditioning regimen are still undetermined for children with LCH. This review aimed to provide an update on advances in allo-HSCT for LCH in children, including indications, stem cell sources, conditioning regimens, chimerism, transplant-related complications, outcomes, and treatment of relapse.

Differentiation of stem cells into chondrocytes and their potential clinical application in cartilage regeneration.

Cartilage diseases and injuries are considered difficult to treat owing to the low regenerative capacity of this tissue. Using stem cells (SCs) is one of the potential methods of treating cartilage defects and creating functional cartilage models for transplants. Their ability to proliferate and to generate functional chondrocytes, a natural tissue environment, and extracellular cartilage matrix, makes SCs a new opportunity for patients with articular injuries or incurable diseases, such as osteoarthritis (OA). The review summarizes the most important scientific reports on biology and mechanisms of SC-derived chondrogenesis and sources of SCs for chondrogenic purposes. Additionally, it focuses on the genetic mechanisms, microRNA (miRNA) regulation, and epigenetic processes steering the chondrogenic differentiation of SCs. It also describes the attempts to create functional cartilage with tissue engineering using growth factors and scaffolds. Finally, it presents the challenges that researchers will have to face in the future to effectuate SC differentiation methods into clinical practice for treating cartilage diseases.

Stem cell-derived exosome treatment for acute spinal cord injury: a systematic review and meta-analysis based on preclinical evidence

BackgroundThe study aims were to systematically review and analyze preclinical research on the efficacy of exosomes derived from various mesenchymal stem cell sources (MSC-exos) for the treatment of spinal cord contusion injury (SCI) in small animal models.MethodsWe conducted a systematic search of PubMed, Embase and Google Scholar databases from their inception through February 29, 2024, to identify eligible English-language studies based on predefined inclusion and exclusion criteria. Two independent investigators performed literature screening, data extraction and bias assessment.ResultsA total of 235 rats were used to assess locomotor recovery at the initial assessment, and exhibited significant improvement in hind limb movement in those treated with exosomes, as indicated by a statistically significant increase in Basso-Beattie-Bresnahan (BBB) scores (MD: 1.26, 95% CI: 1.14–1.38, p &amp;lt; 0.01) compared to the controls. This trend persisted in final assessment data across 21 studies, with pooled analysis confirming similar results (MD: 1.56, 95% CI: 1.43–1.68, p &amp;lt; 0.01). Funnel plot analysis indicated asymmetry in the pooled BBB scores at both baseline and endpoint assessments, suggesting potential publication bias. Exosomes were derived from bone marrow, adipose tissue, umbilical cord or human placental MSCs. Meta-analysis results showed no statistically significant differences in therapeutic efficacy among these MSC-exos sources at various treatment time points.ConclusionMSC-exos demonstrated considerable promise in improving motor function in SCI-affected rats, with bone marrow MSC-derived exosomes having particularly notable effectiveness.

Bone Marrow Mononuclear Cell Transplantation Promotes Bone Healing via Gap Junction-Mediated Cell-Cell Interaction.

Bone marrow mononuclear cells (BM-MNCs) are a rich source of hematopoietic stem cells that have been widely used in experimental therapies for patients with various diseases, including fractures.Activation of angiogenesis is believed to be one of the major modes of action of BM-MNCs; however, the essential mechanism by which BM-MNCs activate angiogenesis remains elusive. This study aimed to demonstrate that BM-MNCs promote bone healing by enhancing angiogenesis through direct cell-to-cell interactions via gap junctions, in addition to a previously reported method. Using a murine fracture model, we aimed to elucidate the relationship between gap junction-mediated cell-to-cell interactions and enhanced fracture healing after BM-MNC transplantation. We evaluated the transfer of substances from BM-MNCs to vascular endothelial cells and osteoblasts in the tissues surrounding the fracture site and assessed the effects of BM-MNC transplantation on bone healing, angiogenesis, and osteogenesis. BM-MNCs transferred substances to vascular endothelial cells and osteoblasts in the tissues surrounding the fracture site. Moreover, BM-MNC transplantation promoted bone healing via gap junction-mediated cell-to-cell interactions, accelerating both angiogenesis and osteogenesis. Our findings provide a novel understanding of fracture healing mechanisms and suggest that BM-MNC transplantation enhances bone healing through gap junction-mediated cell-to-cell interactions, contributing to the development of regenerative medicine strategies targeting bone repair.

Graft ischemia post cell transplantation to the brain: Glucose deprivation as the primary driver of rapid cell death.

Replacing cells lost during the progression of neurodegenerative disorders holds potential as a therapeutic strategy. Unfortunately, the majority of cells die post-transplantation, which creates logistical and biological challenges for cell therapy approaches. The cause of cell death is likely to be multifactorial in nature but has previously been correlated with hypoxia in the graft core. Here we use mathematical modelling to highlight that grafted cells experiencing hypoxia will also face a rapid decline in glucose availability. Interestingly, three neuron progenitor types derived from stem cell sources, and primary human fetal ventral mesencephalic (VM) cells all remained highly viable in severe hypoxia (0.1​% oxygen), countering the idea of rapid hypoxia-induced death in grafts. However, we demonstrate that glucose deprivation, not a paucity of oxygen, was a driver of rapid cell death, which was compounded in ischemic conditions of both oxygen and glucose deprivation. Supplementation of glucose to rat embryonic VM cells transplanted to the adult rat brain failed to improve survival at the dose administered and highlighted the problems of using osmotic minipumps in assisting neural grafting. The data shows that maintaining sufficient glucose in grafts is likely to be of critical importance for cell survival, but better means of achieving sustained glucose delivery is required.

Single-cell Technology in Stem Cell Research.

Single-cell technology (SCT), which enables the examination of the fundamental units comprising biological organs, tissues, and cells, has emerged as a powerful tool, particularly in the field of biology, with a profound impact on stem cell research. This innovative technology opens new pathways for acquiring cell-specific data and gaining insights into the molecular pathways governing organ function and biology. SCT is not only frequently used to explore rare and diverse cell types, including stem cells, but it also unveils the intricacies of cellular diversity and dynamics. This perspective, crucial for advancing stem cell research, facilitates non-invasive analyses of molecular dynamics and cellular functions over time. Despite numerous investigations into potential stem cell therapies for genetic disorders, degenerative conditions, and severe injuries, the number of approved stem cell-based treatments remains limited. This limitation is attributed to the various heterogeneities present among stem cell sources, hindering their widespread clinical utilization. Furthermore, stem cell research is intimately connected with cutting-edge technologies, such as microfluidic organoids, CRISPR technology, and cell/tissue engineering. Each strategy developed to overcome the constraints of stem cell research has the potential to significantly impact advanced stem cell therapies. Drawing from the advantages and progress achieved through SCT-based approaches, this study aims to provide an overview of the advancements and concepts associated with the utilization of SCT in stem cell research and its related fields.

LHPL as Substitute for FBS in Stem Cell Culture by FC

With the advent of regenerative medicine, new methods for treating many incurable diseases might be used to replace damaged, dead, or diseased cells using cell-based methodologies. Mesenchymal stem cells possess immunomodulatory, tissue-protective, and pro-angiogenic properties that make them highly promising for cell-based therapies. Umbilical cord tissue (UCT) is the most common source of mesenchymal stem cells (MSCs) used for explant culture because of its potential for ex-vivo proliferation. Fetal bovine serum (FBS) is the conventional and promising present medium used to culture the MSCs. It is derived from the blood of cow fetus on cardiac puncture without anaesthesia that causes a lot of pain and discomfort to the fetus. This makes its availability limited for research work. Also using a xenogeneic supplement like a foetal bovine serum (FBS) to grow MSCs outside the body increases the risk of zoonotic infection transmission and unfavourable immunological reactions. The self-life of MHC is also very limited. A lot of research is going on to find an alternate to FBS of the above reasons like, human serum, adult bovine serum, lyophilized human platelet lysate (LHPL and chemical formulations. Our research here experimented the potency of lyophilized human platelet lysate (LHPL) as an alternate for FBS in culturing MSCs by using flow cytometry and proved, LHPL is a non-xenogeneic growth supplement with a longer self-life that will sustain the development of MSCs effective and can be stored and reconstituted on demand.

Lineage tracing studies suggest that the placenta is not a de novo source of hematopoietic stem cells.

Definitive hematopoietic stem and progenitor cells (HSPCs) arise from a small number of hemogenic endothelial cells (HECs) within the developing embryo. Understanding the origin and ontogeny of HSPCs is of considerable interest and potential therapeutic value. It has been proposed that the murine placenta contains HECs that differentiate into HSPCs. However, during human gestation HSPCs arise in the aorta considerably earlier than when they can first be detected in the placenta, suggesting that the placenta may primarily serve as a niche. We found that the Runx1 transcription factor, which is required to generate HSPCs from HECs, is not expressed by mouse placental ECs. To definitively determine whether the mouse placenta is a site of HSPC emergence, we performed lineage tracing experiments with a Hoxa13Cre allele that specifically labels ECs in the placenta and umbilical cord (UC), but not in the yolk sac or embryo. Immunostaining revealed Hoxa13Cre lineage-traced HECs and HSPCs in the UC, a known site of HECs, but not the placenta. Consistent with these findings, ECs harvested from the E10.5 aorta and UC, but not the placenta, gave rise to hematopoietic cells ex vivo, while colony forming assays using E14.5 fetal liver revealed only 2% of HSPCs arose from Hoxa13-expressing precursors. In contrast, the pan-EC Cdh5-CreERT2 allele labeled most HSPCs in the mouse placenta. Lastly, we found that RUNX1 and other HEC genes were not expressed in first-trimester human placenta villous ECs, suggesting that human placenta is not hemogenic. Our findings demonstrate that the placenta functions as a site for expansion of HSPCs that arise within the embryo proper and is not a primary site of HSPC emergence.

Osteogenic Differentiation of Adipose Tissue-Derived Mesenchymal Stem Cells on Composite Polymeric Scaffolds: A Review.

The mesenchymal stem cells (MSCs) are the fundamental part of bone tissue engineering for the emergence of reconstructive medicine. Bone tissue engineering has recently been considered a promising strategy for treating bone diseases and disorders. The technique needs a scaffold to provide an environment for cell attachment to maintain cell function and a rich source of stem cells combined with appropriate growth factors. MSCs can be isolated from adipose tissue (ASCs), bone marrow (BM-MSCs), or umbilical cord (UC-MSCs). In the present study, the potential of ASCs to stimulate bone formation in composite polymeric scaffolds was discussed and it showed that ASCs have osteogenic ability in vitro. The results also indicated that the ASCs have the potential for rapid growth, easier adipose tissue harvesting with fewer donor site complications and high proliferative capacity. The osteogenic differentiation capacity of ASCs varies due to the culture medium and the addition of factors that can change signaling pathways to increase bone differentiation. Furthermore, gene expression analysis has a significant impact on improving our understanding of the molecular pathways involved in ASCs and, thus, osteogenic differentiation. Adding some drugs, such as dexamethasone, to the biomaterial composite also increases the formation of osteocytes. Combining ASCs with scaffolds synthesized from natural and synthetic polymers seems to be an effective strategy for bone regeneration. Applying exopolysaccharides, such as schizophyllan, chitosan, gelatin, and alginate in composite scaffolds enhances the osteogenesis potential of ASCs in bone tissue regeneration.

Transplantation of derivative retinal organoids from chemically induced pluripotent stem cells restored visual function

As an emerging type of pluripotent stem cells, chemically induced pluripotent stem cells (CiPSCs) avoid the risks of genomic disintegration by exogenous DNAs from viruses or plasmids, providing a safer stem cell source. To verify CiPSCs’ capacity to differentiate into retinal organoids (ROs), we induced CiPSCs from mouse embryonic fibroblasts by defined small-molecule compounds and successfully differentiated the CiPSCs into three-dimensional ROs, in which all major retinal cell types and retinal genes were in concordance with those in vivo. We transplanted retinal photoreceptors from ROs into the subretinal space of retinal degeneration mouse models and the cells could integrate into the host retina, establish synaptic connections, and significantly improve the visual functions of the murine models. This proof-of-concept study for the first time demonstrated that CiPSCs could differentiate into ROs with a full spectrum of retinal cell types, and provided new insights into chemical approach-based retinal regeneration for degenerative diseases.

Innovations in Epidermal Tissue Engineering

This paper reviews recent advances and challenges in epidermal medical engineering and scaffold material research. The basic elements of tissue engineering include seed cells, scaffold materials and bioactive factors. Current skin tissue engineering techniques offer innovative therapeutic avenues for the repair of large wounds, mainly through suitable tissue scaffolds. The paper also discusses the principles of skin regeneration, including skin structure and function, the process of skin regeneration, and explores the potential of stem cells and exosomes in scar-free healing. Future research directions will focus on developing high-performance biomaterials and scaffolds, optimising stem cell delivery methods, finding optimal stem cell sources and maintaining stem cell activity. In particular, the combined use of natural polymers such as filipin proteins and sodium alginate is expected to create composites that more closely resemble extracellular matrices, thus better promoting cell proliferation and differentiation. In addition, the study of stem cells and exosomes offers new ways to achieve scar-free healing, potentially improving healing efficiency and reducing scar formation when combined with bioscaffolds.

Resolving the design principles that control postnatal vascular growth and scaling.

After birth, tissues grow continuously until reaching adult size, with each organ exhibiting unique cellular dynamics, growth patterns, and (stem or non-stem) cell sources. Using a suite of experimental and computational multiscale approaches, we found that aortic expansion is guided by specific biological principles and scales with the vertebral column rather than animal body weight. Expansion proceeds via two distinct waves of arterial cell proliferation along blood flow that are spatially stochastic, yet temporally coordinated. Each wave exhibits unique cell cycle kinetics and properties, with the first wave exhibiting cell cycle durations as fast as 6 hours. Single-cell RNA sequencing showed changes in fatty acid metabolism concomitant with an increase in cell size. Mathematical modeling and experiments indicated endothelial cell extrusion is essential for homeostatic aortic growth and balancing excess proliferation. In a genetic model of achondroplasia, the aorta achieves proper scaling through enhanced cell extrusion while maintaining normal proliferation dynamics. Collectively, these results provide a blueprint of the principles that orchestrate aortic growth which depends entirely on differentiated cell proliferation rather than resident stem cells.

Chondrogenic commitment of human umbilical cord blood and umbilical cord-derived mesenchymal stem cells induced by the supernatant of chondrocytes: A comparison study.

Native cartilage has low capacity for regeneration because it has very few progenitor cells. Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) and human umbilical cord-derived MSCs (hUC-MSCs) have been employed as promising sources of stem cells for cartilage injury repair. Reproduction of hyaline cartilage from MSCs remains a challenging endeavor. The paracrine factors secreted by chondrocytes possess the capability to induce chondrogenesis from MSCs. The conditioned medium derived from chondrocytes was utilized to induce chondrogenic differentiation of hUCB-MSCs and hUC-MSCs. The expression levels of collagen type I alpha 1 chain (Col1a1), collagen type II alpha 1 chain (Col2a1), and SRY-box transcription factor 9 (SOX9) were assessed through quantitative real-time polymerase chain reaction (qRT-PCR), Western blot (WB), and immunofluorescence (IF) assays. To elucidate the mechanism of differentiation, the concentration of transforming growth factor-β1 (TGF-β1) in the conditioned medium of chondrocytes was quantified using enzyme-linked immunosorbent assay (ELISA). Meanwhile, the viability of cells was assessed using Cell Counting Kit-8 (CCK-8) assays. The expression levels of Col2a1 and SOX9 were found to be higher in induced hUC-MSCs compared to those in induced hUCB-MSCs. The conditioned medium of chondrocytes contained TGF-β1. The CCK-8 assays revealed that the proliferation rate of hUC-MSCs was significantly higher compared to that of hUCB-MSCs. The chondrogenic potential and proliferation capacity of hUC-MSCs surpass those of hUCB-MSCs, thereby establishing hUC-MSCs as a superior source of seed cells for cartilage tissue engineering.

Development of molecular sterility assay for rapid quality release of cord blood erythrocytes units for transfusion.

Umbilical cord blood (CB) units stored in banks are an important source of hematopoietic stem cells for transplantation and other cell therapies. New applications, such as their use in transfusions, require rapid quality release as cord blood red blood cells (CB-RBC) have a shorter shelf life. This project aims to investigate the most prevalent microbial contaminants in CB preparations and validate a rapid sterility testing strategy for CB-RBC based on an automated system (BACT/ALERT®) in tandem with a molecular assay (real-time PCR) capable of detecting at least 100 CFU/mL of Cutibacterium acnes in CB-RBC to accelerate the detection of the most common slow-growing bacteria. Microbial contamination incidence was assessed by reviewing 4696 CB sterility tests, revealing a positivity rate of 3.4%, with C. acnes being the most common slow-growing pathogen. The BACT/ALERT® system, which was validated according to European Pharmacopeia guidelines, was an appropriate method for sterility testing of CB-RBC, although it required up to 14 days of culture to detect C. acnes when iFAPlus and iFNPlus bottles were used to neutralize antimicrobials. Interestingly, the BACT/ALERT® method detected C. acnes at 30 CFU/mL within 14 days, while real-time PCR identified concentrations ≥65 CFU/mL by Day 4. In conclusion, we developed a rapid sterility testing strategy that combines automated culture systems and real-time PCR for early microbial contamination, enhancing CB-RBC shelf life for transfusion and emphasizing the importance of combining detection methods.

The Chondrogenic Potential of the Bovine Tendon Sheath - a Novel Source of Stem Cells for Cartilage Repair.

The human hand is traumatized more frequently than any other bodily part. Trauma and pathological processes (e.g., rheumatoid arthritis, osteoarthritis) commonly implicate the finger joints and specifically damage also the layer of articular cartilage. Endeavors are now being made to surgically repair such cartilage lesions biologically using tissue-engineering approaches that draw on donor cells and/or donor tissues. The tendon sheaths, particularly their inner layers, i.e., the peritendineum, surround the numerous tendons in the hand. The peritendineum is composed of mesenchymal tissue. We hypothesize that this tissue harbors pluripotent mesenchymal stem cells and thus could be used for cartilage repair, irrespective of the donor's age. Using a bovine model (young calves vs. adult cows), the pluripotentiality of the peritendineal stem cells, namely, their osteogenicity, chondrogenicity, and adipogenicity, was investigated by implementing conventional techniques. Subsequently, the chondrogenic potential of the peritendineal tissue itself was analyzed. Its differentiation into cartilage was induced by the application of specific growth factors (members of the TGF-β-superfamily). The characteristics of the tissue formed were evaluated structurally (immuno) histochemically, histomorphometrically, and biochemically (gene expression and protein level). Our data confirm that the bovine peritendineum contains stem cells whose pluripotentiality is independent of donor age. This tissue could also be induced to differentiate into cartilage, likewise, irrespective of the donor's age. Preliminary investigations with adult human peritendineal biopsy material derived from the hand's peritendineal flexor tendon sheaths revealed that this tissue can also be induced to differentiate into cartilage.

Posterior iliac crest vs. proximal tibia: distinct sources of anti-inflammatory and regenerative cells with comparable 6-month clinical outcomes in treatment of osteoarthritis

BackgroundHuman bone marrow is a source of mesenchymal stem cells (MSCs), other progenitor cells, and factors with anti-inflammatory and regenerative capacity. Though the fraction of MSCs out of the nucleated cells is very small, bone marrow aspirate (BMA) for osteoarthritis (OA) has noteworthy effects. BMA is usually collected from the posterior or anterior iliac crest, and rarely from the proximal tibia. We investigated the clinically beneficial concentration of ex vivo MSCs, derived from BM harvested from the posterior iliac crest and proximal tibia by Marrow Cellution™ Aspiration System, and their phenotypic differences, in comparison to autologous Platelet-Rich Plasma (PRP) treatment prepared with a manual, closed system.MethodsA single-center, parallel, randomized controlled study was designed to investigate the efficacy of BMA from the posterior iliac crest compared to BMA from the proximal tibia, against a control group treated with PRP, in knee OA. Thirty patients with knee OA grade I-IV, according to Kellgren-Lawrence (KL), were distributed into each group. Visual Analog Scale (VAS) and Western Ontario & McMaster Universities Arthritis Index (WOMAC) score were used for clinical outcome evaluation.ResultsData from an intermediate analysis of 6-months follow-up, involving 15 patients in each arm, showed that the posterior iliac crest was significantly more densely populated with mononuclear cells, than the proximal tibia (p = 0.005). Flow cytometric analysis on ex vivo BMA showed a significantly greater number of MSCs in the BM-derived from the posterior iliac crest when compared with the proximal tibia (p < 0.001), together with a significantly higher number of platelets (PLTs) (p < 0.001). Surprisingly, despite these differences in cells number, the improvement in early pain and function scores, after each treatment, were statistically significant within each of the three arms. BM from the proximal tibia showed the highest ΔWOMAC, while BM from the posterior iliac crest showed the highest ΔVAS; however, these differences were not statistically significant across the three arms (p > 0.05). A better outcome, in terms of ΔVAS, was observed in patients classified as KL I-II, when treated with BMA from crest (p < 0.001) and PRP (p = 0.004). Moreover, the effect of BMA treatment on ΔVAS depends on MSCs % only in the Tibia Arm (r = -0.59, p = 0.021), where we also found a correlation between ΔWOMAC and monocytes (r = 0.75, p = 0.016).ConclusionThe results indicate that the iliac crest yields a higher concentration of MSCs compared to the proximal tibia, however both BM, independently of the MSCs concentration, show a beneficial clinical outcome in the treatment of knee OA. Furthermore, BMA is not superior to PRP treatment.