scholarly journals Quantification and Comprehensive Analysis of Mesenchymal Stromal Cells in Bone Marrow Samples from Sickle Cell Disease Patients with Osteonecrosis

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Tiago O. Ribeiro ◽  
Paula B. Daltro ◽  
Gildasio Cerqueira Daltro ◽  
Songeli M. Freire ◽  
Roberto Meyer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Cell Number ◽  
Cell Phenotype ◽  
Cell Disease ◽  
Age Related

The potential use of bone marrow mesenchymal stromal cells (BM-MSCs) for the treatment of osteonecrosis in sickle cell disease (SCD) patients is increasing. However, convenient BM-MSC quantification and functional property assays are critical factors for cell-based therapies yet to be optimized. This study was designed to quantify the MSC population in bone marrow (BM) samples from SCD patients with osteonecrosis (SCD group) and patients with osteoarticular complications not related to SCD (NS group), using flow cytometry for CD271+CD45-/low cell phenotype and CFU-F assay. We also compared expanded BM-MSC osteogenic differentiation, migration, and cytokine secretion potential between these groups. The mean total cell number, CFU-F count, and CD271+CD45-/low cells in BM mononuclear concentrate were significantly higher in SCD than in NS patients. A significant correlation between CD271+CD45-/low cell number and CFU-F counts was found in SCD ( r = 0.7483 ; p = 0.0070 ) and NS ( r = 0.7167 ; p = 0.0370 ) BM concentrates. An age-related quantitative reduction of CFU-F counts and CD271+CD45-/low cell number was noted. Furthermore, no significant differences in the morphology, replicative capacity, expression of surface markers, multidifferentiation potential, and secretion of cytokines were found in expanded BM-MSCs from SCD and NS groups after in vitro culturing. Collectively, this work provides important data for the suitable measurement and expansion of BM-MSC in support to advanced cell-based therapies for SCD patients with osteonecrosis.

scholarly journals Bone Marrow Derived Mesenchymal Stromal Cells from Patients with Sickle Cell Disease Display Normal Phenotype and Immunomodulatory Capacity

2016 ◽  
Vol 22 (3) ◽  
pp. S148-S149
Author(s):  
Elizabeth Stenger ◽  
Raghavan Chinnadurai ◽  
Shala Yuan ◽  
Marco Garcia ◽  
Lakshmanan Krishnamurti ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Cell Disease ◽  
Normal Phenotype ◽  
Immunomodulatory Capacity

Murine bone marrow mesenchymal stromal cells have reduced hematopoietic maintenance ability in sickle cell disease

Blood ◽  
2021 ◽  
Author(s):  
Alice Tang ◽  
Ana Nicolle Strat ◽  
Mahmudur Rahman ◽  
Helen Zhang ◽  
Weili Bao ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Bone Marrow ◽  
Stem Cell ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Tissue Injury ◽  
Cell Disease ◽  
Hematopoietic Stem

Sickle Cell Disease (SCD) is characterized by hemolytic anemia, which can trigger oxidative stress, inflammation, and tissue injury that contributes to disease complications. Bone marrow mesenchymal stromal cells (MSCs) tightly regulate hematopoietic stem cell (HSC) homeostasis in health and disease but their functionality in SCD remains unclear. We identified for the first time murine SCD MSCs to have altered gene signatures, reduced stem cell properties, and increased oxidative stress, due in part to hemolysis. Murine SCD MSCs had lower HSC maintenance ability in vitro and in vivo as manifested by increased HSC mobilization and decreased HSC engraftment following transplant. Activation of TLR4 through p65 in MSCs further contributed to MSC dysfunction. Transfusions led to improved MSC and HSC oxidative state in SCD mice. Improving the regulation between MSCs and HSCs has vital implications for enhancing clinical HSC transplantation and gene therapy outcomes and for identification of new molecular targets for alleviating SCD complications.

scholarly journals Bone Marrow–Derived Mesenchymal Stromal Cells from Patients with Sickle Cell Disease Display Intact Functionality

2017 ◽  
Vol 23 (5) ◽  
pp. 736-745 ◽  
Author(s):  
Elizabeth O. Stenger ◽  
Raghavan Chinnadurai ◽  
Shala Yuan ◽  
Marco Garcia ◽  
Dalia Arafat ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Cell Disease

Intact phenotype and functionality of bone marrow derived mesenchymal stromal cells from individuals with sickle cell disease

Cytotherapy ◽  
2017 ◽  
Vol 19 (5) ◽  
pp. S45
Author(s):  
E. Stenger ◽  
R. Chinnadurai ◽  
S. Yuan ◽  
M. Garcia ◽  
D. Arafat ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Cell Disease

Apoliprotein A-I Mimetic Peptide and Sickle Vasculopathy: Mouse Model Study of Acute Administration.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1521-1521
Author(s):  
Lewis L. Hsu ◽  
Valeriani Bead ◽  
Lita A. Freeman ◽  
Gregory J. Kato ◽  
James G Taylor ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Bone Marrow ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Acute Administration ◽  
Wild Type ◽  
Cell Disease ◽  
Mimetic Peptide ◽  
Mimetic Peptides

Abstract Abstract 1521 Poster Board I-544 Epidemiologic observations associate low levels of Apolipoprotein A-1 (ApoA-I) and low high density lipoprotein (HDL) with greater prevalence of pulmonary hypertension in sickle cell patients. Sickle cell mice with knockout of apoliproteins have greater pulmonary hypertension and endothelial dysfunction than sickle cell mice with wild-type levels of ApoA-I, and they have further worsening of the global dysregulation of the nitric oxide axis (NO) seen in sickle vasculopathy. Ou et al. (Circulation 2003; 107(18):2337-41) showed that chronic administration (3-4 weeks) of the ApoA-I mimetic L-4F improves vasodilation in hypercholesterolemic and sickle mice. Remaley et al. (Circulation 2006;114:II_23) showed atheroprotective effects after a single (1mg/mouse) intravenous administration of the ApoA-I mimetic peptide 5A as a result of increased reverse cholesterol transfer. These preclinical data provide a mechanistic basis for the effects of ApoA-I in sickle cell disease and suggest that ApoA-I mimetics deserve further study as a potential therapy for sickle cell disease. However, it is unknown whether acute administration of an ApoA-I mimetic has any efficacy for sickle cell disease, and whether all ApoA-I mimetic peptides will be helpful. We hypothesized that acute administration of ApoA-I mimetic could improve endothelial dysfunction in sickle cell mouse models. Bone marrow harvested from Berkeley sickle cell mouse donors and hemizygote non-sickling control mice were used to generate three groups of mice after myeloablative irradiation of wild-type C57BL6 recipient mice. Three million cells whole marrow was injected per mouse. Mice were fully engrafted by 3 months. Hemoglobin analysis showed one group produced only sickle RBC, another group only hemizygote RBC, and a third group were mixed chimeras with 30% sickle RBC and 70% hemizygote RBC (intended to model a sickle cell patient on chronic transfusion). Pulse-wave velocities, a measure of arterial stiffness, showed no improvement after IV injection of the ApoA-I mimetic peptide, 5A-POPC (170 mcl of 8.6 mg/ml). Aortic rings prepared from mice of all three groups showed blunted relaxation response to acetylcholine and sodium nitroprusside, as expected for abnormal nitric oxide bioavailability - these vasorelaxation responses were not significantly different in the absence vs. presence of 5A-POPC incubation in vitro. These results agree with our previous observation that sickle bone marrow transplanted into transgenic mice overexpressing ApoA-I had no difference in vasculopathy from sickle bone marrow transplanted into wild-type mice We conclude that acute exposure to this ApoA-I mimetic, either in vivo or in vitro, is not sufficient to relieve the vasculopathy of sickle cell mouse models. Higher doses or long term therapy may be necessary for benefit. An alternative explanation is that ApoA-I mimetic peptides differ in their benefit for sickle cell vasculopathy, and that the key property is the high level of oxidant scavenging by L-4F rather than cholesterol removal by 5A-POPC. Future studies can determine whether the cardiovascular benefits of ApoA-I mimetics correlate with their antioxidant function. Disclosures No relevant conflicts of interest to declare.

scholarly journals Kinship of conditionally immortalized cells derived from fetal bone to human bone-derived mesenchymal stroma cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
S. Marozin ◽  
B. Simon-Nobbe ◽  
S. Irausek ◽  
L. W. K. Chung ◽  
G. Lepperdinger
Keyword(s):  
Bone Marrow ◽  
Cell Line ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Human Bone ◽  
Multilineage Differentiation ◽  
Differentiation Potential ◽  
Osteoblast Cell Line ◽  
Age Related

AbstractThe human fetal osteoblast cell line (hFOB 1.19) has been proposed as an accessible experimental model for study of osteoblast biology relating to drug development and biomaterial engineering. For their multilineage differentiation potential, hFOB has been compared to human mesenchymal progenitor cells and used to investigate bone-metabolism in vitro. Hereby, we studied whether and to what extent the conditionally immortalized cell line hFOB 1.19 can serve as a surrogate model for bone-marrow derived mesenchymal stromal cells (bmMSC). hFOB indeed exhibit specific characteristics reminiscent of bmMSC, as colony formation, migration capacity and the propensity to grow as multicellular aggregates. After prolonged culture, in contrast to the expected effect of immortalization, hFOB acquired a delayed growth rate. In close resemblance to bmMSC at increasing passages, also hFOB showed morphological abnormalities, enlargement and finally reduced proliferation rates together with enhanced expression of the cell cycle inhibitors p21 and p16. hFOB not only have the ability to undergo multilineage differentiation but portray several important aspects of human bone marrow mesenchymal stromal cells. Superior to primary MSC and osteoblasts, hFOB enabled the generation of continuous cell lines. These provide an advanced basis for investigating age-related dysfunctions of MSCs in an in vitro 3D-stem cell microenvironment.

Abstract 607: Kininogen Regulates Thrombin Generation in a Mouse Model of Sickle Cell Disease

2015 ◽  
Vol 35 (suppl_1) ◽  
Author(s):  
Erica M Sparkenbaugh ◽  
Kasemsiri Chandarajoti ◽  
Nigel S Key ◽  
Andras Gruber ◽  
Nigel Mackman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Thrombin Generation ◽  
Vascular Inflammation ◽  
Factor Xa ◽  
Cell Disease ◽  
Intrinsic Pathway ◽  
Coagulation Pathway

Sickle cell disease (SCD) is associated with activation of coagulation and vascular inflammation. We previously demonstrated that, in mouse models of SCD, tissue factor (TF) expressed on leukocytes activates coagulation and contributes to inflammation via microvascular thrombosis, whereas non-coagulant form of TF expressed by endothelial cells mediates factor Xa (FXa)-PAR2 signalling and contributes to inflammation via IL-6 expression. We also showed that both rivaroxaban and dabigatran, oral inhibitors of FXa and thrombin, attenuated the hypercoagulable state in sickle mice. It has been proposed that anticoagulants that target the intrinsic coagulation pathway may be associated with a lower risk of bleeding compared to targeting the extrinsic or common coagulation pathways. Therefore, we tested if the intrinsic pathway contributes to the activation of coagulation in sickle cell mice. Bone marrow from sickle (SS) and non-sickle (AA) Townes mice was transplanted into wild-type (WT) (n=18 AA, 15 SS), FXII-/- (n=14 AA, 22 SS), or FXI-/- (n=21 AA, 23 SS) recipients (all on C57Bl/6 genetic background). In addition, bone marrow from AA or SS mice was transplanted into high molecular weight kininogen (HK)+/+ (n=17 AA, 16 SS) or HK-/- (n=10 AA, 19 SS) mice. All mice were used for experiments 5 months after BMT. Thrombin generation, measured by plasma thrombin anti-thrombin (TAT) levels, was increased in WT mice injected with SS bone marrow (WT/BMSS) compared to WT/BMAA mice (4.4±0.7 vs 2.6±0.3 ng/mL, p<0.05, mean ± S.E.M), however neither FXII nor FXI deficiency affected this parameter. Consistent with these data, inhibition of FXIIa-dependent activation of FXI with 14E11 antibody also did not reduce plasma TAT levels in SS Townes mice. Interestingly, elevated plasma TAT levels observed in HK+/+/BMSS mice was significantly reduced in HK-/-/BMSS mice (4.8±0.5 versus 3.48±0.2 ng/mL, p<0.05). These data indicate that HK, but not FXII and FXI, contributes to thrombin generation in SCD at steady state of disease. In vitro, HK fragments induce TF expression on monocytes via activation of CD11b/18. We are now investigating if inhibition of HK fragments-induced TF expression on monocytes may attenuate the hypercoagulabIe state in SCD mice without impacting hemostasis.

scholarly journals Autologous bone marrow stromal cells are promising candidates for cell therapy approaches to treat bone degeneration in sickle cell disease

2015 ◽  
Vol 15 (3) ◽  
pp. 584-594 ◽  
Author(s):  
Angélique Lebouvier ◽  
Alexandre Poignard ◽  
Laura Coquelin-Salsac ◽  
Julie Léotot ◽  
Yasuhiro Homma ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Sickle Cell Disease ◽  
Cell Therapy ◽  
Sickle Cell ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Autologous Bone Marrow ◽  
Marrow Stromal Cells ◽  
Cell Disease ◽  
Autologous Bone

Autologous mesenchymal stromal cells to support allogeneic hematopoietic stem cell engraftment in sickle cell disease

Cytotherapy ◽  
2015 ◽  
Vol 17 (6) ◽  
pp. S46
Author(s):  
Elizabeth Stenger ◽  
Shala Yuan ◽  
Marco Garcia ◽  
Ian B. Copland ◽  
Jacques Galipeau
Keyword(s):  
Stem Cell ◽  
Sickle Cell Disease ◽  
Hematopoietic Stem Cell ◽  
Sickle Cell ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Cell Disease ◽  
Hematopoietic Stem ◽  
Cell Engraftment

scholarly journals Age-related Changes in Bone Marrow Mesenchymal Stromal Cells

2017 ◽  
Vol 26 (9) ◽  
pp. 1520-1529 ◽  
Author(s):  
Payal Ganguly ◽  
Jehan J. El-Jawhari ◽  
Peter V. Giannoudis ◽  
Agata N. Burska ◽  
Frederique Ponchel ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Current Knowledge ◽  
The Elderly ◽  
Cellular Level ◽  
Age Related ◽  
Population Doublings

Aging at the cellular level is a complex process resulting from accumulation of various damages leading to functional impairment and a reduced quality of life at the level of the organism. With a rise in the elderly population, the worldwide incidence of osteoporosis (OP) and osteoarthritis (OA) has increased in the past few decades. A decline in the number and “fitness” of mesenchymal stromal cells (MSCs) in the bone marrow (BM) niche has been suggested as one of the factors contributing to bone abnormalities in OP and OA. It is well recognized that MSCs in vitro acquire culture-induced aging features such as gradual telomere shortening, increased numbers of senescent cells, and reduced resistance to oxidative stress as a result of serial population doublings. In contrast, there is only limited evidence that human BM-MSCs “age” similarly in vivo. This review compares the various aspects of in vitro and in vivo MSC aging and suggests how our current knowledge on rejuvenating cultured MSCs could be applied to develop future strategies to target altered bone formation processes in OP and OA.

Sign in / Sign up

Export Citation Format

Share Document