The use of mesenchymal stem cells in therapy of steroid-resistance chronic refractory graft-versus-host disease after hematopoietic stem cell transplantation

Introduction. Chronic graft-versus-host disease (GVHD) is a life-threatening complication after allogeneic hematopoietic stem cell transplantation. Mesenchymal stem cells (MSC) are involved in tissue repair and modulating immune responses. We present report of two cases of use of third-party donor mesenchymal stem cells in the therapy of steroid-resistant chronic GVHD. Case reports. The doses of MSC in both patients were 1.08 and 1.4 x 106 cells per kg body weight respectively. Time from diagnosis of GVHD to infusion of MSC was 420 and 330 days, respectively. Both patients had single infusion of MSC. We did not observe any adverse effects after both infusions of MSC. We obtained initially a partial response and then complete response in the first patient and a partial response in the second patient. In the first patient we achieved improvement of skin involvement resulting in reduction of immunosuppression. In the second patient with scleroderma we achieved only a minor improvement. Conclusion. Administration of MSC is a safe method and might by an effective therapy for patients with chronic refractory GVHD.Wstęp. Przewlekła postać choroby przeszczep-przeciwko-gospodarzowi (chronic graft-versus-host disease, GVHD) jest zagrażającym życiu powikłaniem po przesz-czepieniu allogenicznych komórek krwiotwórczych. Mezenchymalne komórki macierzyste (MSC) mają znaczenie w procesach reparacji tkankowej oraz modulowania odpowiedzi immunologicznej. W pracy przedstawiamy opis dwóch przypadków zastosowania MSC od dawcy niezwiązanego z pacjentem, tzw. third-party donor, w terapii steroidoopornej przewlekłej GVHD. Opis przypadku. Zastosowane dawki MSC u pacjentów wynosiły odpowiednio 1,08 oraz 1,4 x 106 komórek na kg cc. Czas od rozpoznania GVHD do infuzji MSC wynosił odpowiednio 420 i 330 dni. U obydwu pacjentów dokonano pojedynczych infuzji MSC. U żadnego z nich nie obserwowano niepożądanych reakcji po zastosowaniu MSC. U pierwszego pacjenta uzyskano początkowo częściową poprawę, a po kilku miesiącach remisję całkowitą wszystkich zmian związanych z GVHD. Dzięki temu możliwe było znaczne zredukowanie dawek leków immunosupresyjnych. U drugiego pacjenta uzyskano tylko przejściową nieznaczną poprawę stanu twardzinowej skóry. Wniosek: Zastosowanie MSC jest metodą bezpieczną i może być efektywną terapią u pacjentów z przewlekłą postacią choroby GVHD

Maternal predictors and quality of umbilical cord blood units

The aim of the study was to determine the relationship between the maternal age at delivery and selected properties of the cord blood stem cells. The study included 50 pregnant women aged between 18 and 38 years in which spontaneous labors or elective cesarean sections were performed. Umbilical cord blood was collected immediately after the women were delivered of newborns. The samples were analyzed in the Polish Stem Cells Bank in Warsaw. The highest mean WBC level (p < 0.05) was observed in the umbilical blood collected from patients aged 35 years and more. Similarly, the highest mean cell viability was observed in the umbilical cord blood collected from patients aged 35 and more. There were no statistically significant correlations between the CD34+ cells count and mean cell viability in the umbilical cord blood and the maternal age. With the significance level at p < 0.001, the females after spontaneous labor revealed a visibly higher WBC level than patients after a cesarean section. The higher mean WBC concentration (24.95 thousand/μl) was observed in the umbilical cord blood of patients aged 35 and more after spontaneous labors. In the same group, the umbilical cord blood was also characterized by the highest mean cell viability (98.72%). The number of nucleated cells in the umbilical cord blood collected in the perinatal period increases together with the maternal age. In the course of physiological spontaneous labors, the collected umbilical cord blood has more nucleated cells as compared with elective caesarian sections

Multilineage differentiation potential of human dental pulp stem cells : impact of 3D and hypoxic environment on osteogenesis in vitro

Human dental pulp harbours unique stem cell population exhibiting mesenchymal stem/stromal cell (MSC) characteristics. This study aimed to analyse the differentiation potential and other essential functional and morphological features of dental pulp stem cells (DPSCs) in comparison with Wharton&rsquo;s jelly-derived MSCs from the umbilical cord (UC-MSCs), and to evaluate the osteogenic differentiation of DPSCs in 3D culture with a hypoxic microenvironment resembling the stem cell niche. Human DPSCs as well as UC-MSCs were isolated from primary human tissues and were subjected to a series of experiments. We established a multiantigenic profile of DPSCs with CD45&minus;/CD14&minus;/CD34&minus;/CD29+/CD44+/CD73+/CD90+/CD105+/Stro-1+/HLA-DR&minus; (using flow cytometry) and confirmed their tri-lineage osteogenic, chondrogenic, and adipogenic differentiation potential (using qRT-PCR and histochemical staining) in comparison with the UC-MSCs. The results also demonstrated the potency of DPSCs to differentiate into osteoblasts in vitro. Moreover, we showed that the DPSCs exhibit limited cardiomyogenic and endothelial differentiation potential. Decreased proliferation and metabolic activity as well as increased osteogenic differentiation of DPSCs in vitro, attributed to 3D cell encapsulation and low oxygen concentration, were also observed. DPSCs exhibiting elevated osteogenic potential may serve as potential candidates for a cell-based product for advanced therapy, particularly for bone repair. Novel tissue engineering approaches combining DPSCs, 3D biomaterial scaffolds, and other stimulating chemical factors may represent innovative strategies for pro-regenerative therapies

Wpływ pH i pCO2 krwi pępowinowej uzyskiwanej okołoporodowo na wybrane parametry komórek macierzystych

Objective: The aim of the study was to demonstrate a correlation between pH and pCO2 levels in umbilical cord blood CD34+ cells and their vitality was analyzed. Material and methods: The study included 50 pregnant women after vaginal delivery at term or elective cesarean section. Umbilical cord blood was collected immediately after birth. The probes were analyzed at the Polish Stem Cell Bank in Warsaw. Results: The number of CD34+ cells ranged from 0.1-0.2 in white blood cells count over 12 thousand/ml and pH of >7.3. If pH ranged between 7.35-7.40, the number of CD34+ was 0.3-0.4. The highest number of CD34+ cells was noted for pH of 7.30-7.35 and amounted to 0.4-0.5. Analysis of stem cell vitality showed that the highest level, over 98%, was obtained when pH was 40.0 mmHg. For pCO2Cel pracy: Celem badania było wykazanie korelacji pomiędzy poziomem pH i pCO2 krwi pępowinowej pobieranej od dawcow, a jakością pozyskanego materiału. Dodatkowo postanowiono znaleźć zależność wpływu pH i pCO2 krwi pępowinowej na ilość komorek CD34+, ich żywotność oraz wpływ sposobu zakończenia ciąży na właściwości krwi pępowinowej. Materiał i metody: Do badania zakwalifikowano 50 ciężarnych kobiet, u ktorych wystąpił porod siłami natury lub elektywne cięcie cesarskie w terminie porodu. Krew pępowinowa została pobrana bezpośrednio po urodzeniu noworodka. Probki poddano analizie w Polskim Banku Komorek Macierzystych w Warszawie. Wyniki: Przy liczbie leukocytow powyżej 12 tys./μl i jednoczesnym pH >7,3 ilość CD34+ mieściła się w przedziale 0,1-0,2. W przypadku gdy pH wynosi 7,35 -7,40 zakres wartości CD34+ wyniosł 0,3-0,4. Największą ilość CD34+ uzyskano dla pH 7,30-7,35 i wynosiła ona 0,4-0,5. Badając żywotność komorek macierzystych, jej największy poziom >98% uzyskano gdy pH wyniosło poniżej 7,3 oraz gdy pH było ≥7,4. Badania wykazały, że żywotność komorek macierzystych obniżała się do 97-98% przy pH 7,3-7,4. Zaobserwowano niskie wartości CD34+ (0,01-0,09) przy wartościach pCO2 >40,0 mmHg. Przy pCO

Impact of graphene-based surfaces on the basic biological properties of human umbilical cord mesenchymal stem cells : implications for ex vivo cell expansion aimed at tissue repair

The potential therapeutic applications of mesenchymal stem/stromal cells (MSCs) and biomaterials have attracted a great amount of interest in the field of biomedical engineering. MSCs are multipotent adult stem cells characterized as cells with specific features, e.g., high differentiation potential, low immunogenicity, immunomodulatory properties, and efficient in vitro expansion ability. Human umbilical cord Wharton&rsquo;s jelly-derived MSCs (hUC-MSCs) are a new, important cell type that may be used for therapeutic purposes, i.e., for autologous and allogeneic transplantations. To improve the therapeutic efficiency of hUC-MSCs, novel biomaterials have been considered for use as scaffolds dedicated to the propagation and differentiation of these cells. Nowadays, some of the most promising materials for tissue engineering include graphene and its derivatives such as graphene oxide (GO) and reduced graphene oxide (rGO). Due to their physicochemical properties, they can be easily modified with biomolecules, which enable their interaction with different types of cells, including MSCs. In this study, we demonstrate the impact of graphene-based substrates (GO, rGO) on the biological properties of hUC-MSCs. The size of the GO flakes and the reduction level of GO have been considered as important factors determining the most favorable surface for hUC-MSCs growth. The obtained results revealed that GO and rGO are suitable scaffolds for hUC-MSCs. hUC-MSCs cultured on: (i) a thin layer of GO and (ii) an rGO surface with a low reduction level demonstrated a viability and proliferation rate comparable to those estimated under standard culture conditions. Interestingly, cell culture on a highly reduced GO substrate resulted in a decreased hUC-MSCs proliferation rate and induced cell apoptosis. Moreover, our analysis demonstrated that hUC-MSCs cultured on all the tested GO and rGO scaffolds showed no alterations of their typical mesenchymal phenotype, regardless of the reduction level and size of the GO flakes. Thus, GO scaffolds and rGO scaffolds with a low reduction level exhibit potential applicability as novel, safe, and biocompatible materials for utilization in regenerative medicine

Fludarabine, treosulfan and etoposide sensitivity and the outcome of hematopoietic stem cell transplantation in childhood acute myeloid leukemia

Background: The prognostic role of the ex vivo drug resistance profile has not yet been proved in childhood acute myeloid leukemia (AML). The aim of the study was the analysis of the impact of the ex vivo drug resistance profile in a cohort of 44 children with AML undergoing hematopoietic stem cell transplantation (HSCT). Patients and Methods: Myeloblasts for drug resistance testing were obtained from the bone marrow either on diagnosis or at relapse, before the HSCT procedure and were tested by the MTT assay. Results: Children who relapsed after transplantation showed higher ex vivo resistance of the leukemic blasts to etoposide, mercaptopurine, thioguanine, fludarabine, mitoxantrone and treosulfan than those who stayed in remission. Despite being nondiscriminative, the combined ex vivo drug resistance profile to fludarabine, treosulfan and etoposide (FTE score) was the strongest prognostic factor by multivariate analysis. Conclusion: The combined drug resistance profile to fludarabine, treosulfan and etoposide may be useful for better stratification of children with AML undergoing stem cell transplantation or to indicate the necessity for additional post-transplant therapy

Cord blood transplantations in Polish pediatric centers: report of the Polish Pediatric Group for Hematopoietic Stem Cell Transplantation

WstępPrzeszczepianie komórek krwiotwórczych krwi pępowinowej (CBT) jest uznaną metodą terapeutyczną, wykonywaną od roku 1988. Pierwsze przeszczepienie w Polsce wyłącznie z krwi pępowinowej wykonano 12 października 2000 r. w Poznaniu.Cel pracyAnaliza wyników przeszczepiania komórek krwiotwórczych w polskich ośrodkach pediatrycznych.Pacjenci i metodykaDo badań włączono 19 pacjentów (5 dziewcząt, 14 chłopców), w wieku 0,1–10 lat (mediana 4,3 roku), u których w latach 2000–2011 w polskich ośrodkach pediatrycznych wykonano przeszczepienie krwi pępowinowej. Pacjentów kierowano do CBT z powodu następujących rozpoznań: ostra białaczka limfoblastyczna (n=6), ostra białaczka mieloblastyczna (n=1), zespół mielodysplastyczny (n=2), zespół Wiskotta i Aldricha (n=3), niedokrwistość Fanconiego (n=2), adrenoleukodystrofia (n=1), histiocytoza Langerhansa (n=1), przewlekła choroba ziarniniakowa (n=1), zespół Kostmanna (n=1), zespół Sandhoffa (n=1). Zastosowano kondycjonowanie mieloablacyjne u 9 pacjentów oraz o zredukowanej toksyczności u 10 pacjentów. Krew pępowinowa pochodziła od dawcy rodzinnego w 8 przypadkach lub od dawcy niespokrewnionego w 11 przypadkach. Zgodność 6/6 HLA wystąpiła w 10 przypadkach.Wyniki10/19 (52,6%) dzieci żyje, mediana przeżycia 3,1 roku (95%CI=1,4–4,7), prawdopodobieństwo przeżycia 2-letniego wynosi 0,409±0,133. Przyczyny zgonów obejmowały brak przyjęcia przeszczepu (n=2), powikłania infekcyjne (n=3) lub wznowę (n=4). Dwoje dzieci z powodu nieprzyjęcia przeszczepu miało wykonane przeszczepienie haploidentyczne. W analizie wielowariantowej, jedynym czynnikiem prognostycznym mającym wpływ na całkowite przeżycie było wystąpienie udokumentowanego uogólnionego zakażenia do dnia +180.WnioskiU pacjentów niemających zgodnego dawcy CBT jest ważną opcją terapeutyczną, dającą szanse wyleczenia dla około połowy pacjentów.BackgroundCord blood transplantation (CBT) is accepted therapeutic method in transplantology since 1988. The first isolated CBT was performed on 12 October 2000 in Poznań.ObjectiveAnalysis of results of CBT in Polish pediatric centers.Patients and methodsA total numer of 19 patients (5 female, 14 male), aged 0.1–10 years (median 4.3yrs) transplanted with cord blood between 2000–2011 in Polish pediatric centers. The initial diagnosis was: acute lymphoblastic leukemia (n=6), acute myeloid leukemia (n=1), myelodysplstic syndrome (n=2), Wiskott-Aldrich syndrome (n=3), Fanconi anemia (n=2), adrenoleukodystrophy (n=1), Langerhans cell histiocytosis (n=1), chronic granulomatous disease (n=1), Kostmann syndrome (n=1), Sandhoff syndrome (n=1). Pre-transplant conditioning was myeloablative in 9 patients and reduced-intensity in 10 patients. The source of cord blood was family donor in 8 cases or unrelated donor in 11 cases. Histocompatibility 6/6 HLA between donor-recipient was present in 10 cases.Results10/19 (52.6%) children stay alive, median survival 3.1 years (95%CI=1.4–4.7), probability of 2-year survival was 0.409±0.133. The cause of death was primary graft failure (n=2), infectious complications (n=3) or relapse (n=4). Two children with primary graft failure had subsequent haploidentical transplantation. In multivariate analysis, generalized documented infection was the only predictive adverse factor of overall survival.ConclusionCBT is an important therapeutic option for patients lacking matched donor, offering positive outcome for a half of patients

Wharton’s Jelly Mesenchymal Stem Cell Administration Improves Quality of Life and Self-Sufficiency in Children with Cerebral Palsy: Results from a Retrospective Study

The aim of this paper was to describe the outcome of the therapeutic administration of allogenic mesenchymal stem cells obtained from Wharton’s jelly (WJ-MSCs) in children with cerebral palsy (CP) during a medical therapeutic experiment. We retrospectively analyzed the records of 109 patients recruited in daily clinical practice. Each patient received 1–10 injections and was examined by the same neurologist (study investigator (SI)) on the day of each infusion. The SI used a 6-point Likert scale to assess the quality of life (QoL) and self-sufficiency of the patients on the basis of the neurological examination. Children with >50% follow-ups after this administration were included into the quantitative analysis. In addition, the assessments of the parents and other health care professionals were obtained for 23 patients and compared with those of the SI. Forty-eight of 54 analyzed patients (88.9%) achieved some improvement in health status. Forty-eight (88.9%) patients experienced an increase in their QoL, and 21 patients (38.9%) achieved an increase in their self-sufficiency level. Improvement was achieved in 17 areas. Adverse events were mild and temporary except one case of epilepsy deterioration leading to treatment discontinuation. Age, body mass, and cell dose were not significant predictors of QoL response, contrary to epilepsy; developmental breakthrough was dose-dependent

Autologous cord blood in children with cerebral palsy: a review

The aim of this narrative review is to report on the current knowledge regarding the clinical use of umbilical cord blood (CB) based on articles from PubMed and clinical trials registered on ClinicalTrials.gov. An increasing amount of evidence suggests that CB may be used for both early diagnostics and treatment of cerebral palsy. The acidity of CB and its biochemical parameters, including dozens of cytokines, growth factors, and other metabolites (such as amino acids, acylcarnitines, phosphatidylcholines, succinate, glycerol, 3-hydroxybutyrate, and O-phosphocholine) are predictors of future neurodevelopment. In addition, several clinical studies confirmed the safety and efficacy of CB administration in both autologous and allogeneic models, including a meta-analysis of five clinical trials involving a total of 328 participants. Currently, nine clinical trials assessing the use of autologous umbilical CB in children diagnosed with hypoxic-ischemic encephalopathy or cerebral palsy are in progress. The total population assessed in these trials exceeds 2500 patients