Proplatelet Formation Dynamics of Mouse Fresh Bone Marrow Explants

10.3791/62501 ◽  
2021 ◽  
Author(s):  
Inès Guinard ◽  
François Lanza ◽  
Christian Gachet ◽  
Catherine Léon ◽  
Anita Eckly
Keyword(s):  
Bone Marrow ◽  
Formation Dynamics ◽  
Proplatelet Formation ◽  
Fresh Bone

Evaluation of new bone formation in irradiated areas using association of mesenchymal stem cells and total fresh bone marrow mixed with calcium phosphate scaffold

2014 ◽  
Vol 25 (12) ◽  
pp. 2711-2720 ◽  
Author(s):  
P. Bléry ◽  
P. Corre ◽  
O. Malard ◽  
S. Sourice ◽  
P. Pilet ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Calcium Phosphate ◽  
Bone Formation ◽  
New Bone Formation ◽  
Fresh Bone

scholarly journals DISTINCT EVENTS IN THE IMMUNE RESPONSE ELICITED BY TRANSFERRED MARROW AND THYMUS CELLS

1969 ◽  
Vol 130 (6) ◽  
pp. 1243-1261 ◽  
Author(s):  
G. M. Shearer ◽  
G. Cudkowicz
Keyword(s):  
Bone Marrow ◽  
Immune Response ◽  
Bone Marrow Cells ◽  
Second Step ◽  
Cell Divisions ◽  
Cellular Events ◽  
Antigen Complex ◽  
Fresh Bone ◽  
Thymus Cells ◽  
Marrow Cells

Marrow cells and thymocytes of unprimed donor mice were transplanted separately into X-irradiated syngeneic hosts, with or without sheep erythrocytes (SRBC). Antigen-dependent changes in number or function of potentially immunocompetent cells were assessed by retransplantation of thymus-derived cells with fresh bone marrow cells and SRBC; of marrow-derived cells with fresh thymocytes and SRBC; and of thymus-derived with marrow-derived cells and SRBC. Plaque-forming cells (PFC) of the direct (IgM) and indirect (IgG) classes were enumerated in spleens of secondary host mice at the time of peak responses. By using this two-step design, it was shown (a) that thymus, but not bone marrow, contained antigen-reactive cells (ARC) capable of initiating the immune response to SRBC (first step), and (b) that the same antigen complex that activated thymic ARC was required for the subsequent interaction between thymus-derived and marrow cells and/or for PFC production (second step). Thymic ARC separated from marrow cells but exposed to SRBC proliferated and generated specific inducer cells. These were the cells that interacted with marrow precursors of PFC to form the elementary units for plaque responses to SRBC, i.e. the class- and specificity-restricted antigen-sensitive units. It was estimated that each ARC generated 80–800 inducer cells in 4 days by way of a minimum of 6–10 cell divisions. On the basis of the available evidence, a simple model was outlined for cellular events in the immune response to SRBC.

scholarly journals The Role of a Combination of Autogenous Fresh Bone Marrow and Omental Free Graft in the Cervical Esophageal Wound Healing in Dogs

2010 ◽  
Vol 5 (3) ◽  
pp. 202-207
Author(s):  
Omid Azari ◽  
Mohammad Mahdi Molaei ◽  
Reza Kheirandis ◽  
Sara Hamzeh Aliabad
Keyword(s):  
Bone Marrow ◽  
Wound Healing ◽  
Free Graft ◽  
Fresh Bone

scholarly journals Photobiomodulation of fresh bone marrow aspirate for regenerative therapy

2021 ◽  
Vol 10 (11) ◽  
pp. e140101119545
Author(s):  
Carolina dos Santos Santinoni ◽  
Liziana Jancos Calles ◽  
Nathália Laís Farias ◽  
Thaís Sanches Leite Patara ◽  
Bianca Eduarda de Lima Neves ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Aspirate ◽  
Positive Control ◽  
Negative Control ◽  
Low Level Laser Therapy ◽  
Adult Rats ◽  
Level Laser ◽  
Group Water ◽  
A Cell ◽  
Fresh Bone

Use of mesenchymal stem cells and low-level laser therapy (LLLT) have been widely studied to promote bone healing. evaluate effect of photobiomodulation on total number of cells (TNC) and cell viability (CV) of fresh bone marrow aspirate (BMA). Femur BMA from 10 adult rats was collected and a cell concentration of 1x107 cell/mL was obtained. Cell suspension was deposited on 96 well cell culture plates and distributed in groups: 1) RPMI, positive control; 2) Distilled Water, negative control; 3) Red Laser (RL); 4) Infrared Laser (IRL). Groups RL and IRL received LLLT application right after incubation. Cells were incubated for 24 h. TNC and CV were assessed through trypan blue assay after 1, 3, 6, 10 and 24 h of incubation. Data distribution was verified by Shapiro-Wilk test. Kruskal-Wallis test was used for intergroup and intragroup comparisons (p<0.05). TNC: after 1 and 3 h, groups RL and IRL presented significantly higher TNC than Group Water; after 6 and 10 h, groups RPMI, RL and IRL presented significantly higher TNC than Group Water. CV: after 1 h, groups RL and IRL showed significantly higher percentage of VC than Group Water; after 3, 6 and 10 h, all groups presented significantly higher percentage of VC than Group Water. It can be concluded that LLLT enhanced number and viability of fresh bone marrow aspirate cells.

The use of bone-marrow-derived fibroblastoid cells and fresh bone marrow in the treatment of bone defects: an experimental study

1997 ◽  
Vol 26 (1) ◽  
pp. 55-60 ◽  
Author(s):  
G. Krzymanski ◽  
M. Kalczak ◽  
W. Wiktor-Jedrzejczak
Keyword(s):  
Bone Marrow ◽  
Experimental Study ◽  
Bone Defects ◽  
Fresh Bone

scholarly journals The effect of fresh bone marrow cells on reconstruction of mouse calvarial defect combined with calvarial osteoprogenitor cells and collagen-apatite scaffold

2012 ◽  
Vol 7 (12) ◽  
pp. 974-983 ◽  
Author(s):  
Xiaohua Yu ◽  
Liping Wang ◽  
Fei Peng ◽  
Xi Jiang ◽  
Zengmin Xia ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Calvarial Defect ◽  
Osteoprogenitor Cells ◽  
Fresh Bone ◽  
Marrow Cells

Ponatinib Induced Thrombocytopenia Might Inhibit Proplatelet Formation of Megakaryocytes Via the Pathways Including Rho/Rock and Rac.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2186-2186
Author(s):  
Kazunori Murai ◽  
Shugo Kowata ◽  
Tatsuo Oyake ◽  
Tadashi Shimoyama ◽  
Maki Asahi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Vitro Culture ◽  
Bone Marrow Cells ◽  
Molecular Response ◽  
Severe Thrombocytopenia ◽  
Western Blots ◽  
T315i Mutation ◽  
Proplatelet Formation ◽  
Marrow Cells

Abstract Abstract 2186 Background: Ponatinib (AP24534) is identified as a pan-BCR-ABL inhibitor that potently inhibits the T315I gatekeeper mutant, and has advanced into clinical development for the treatment of refractory or resistant CML (Chronic Myeloid Leukemia). Ponatinib potently inhibited in vitro proliferation of Ba/F3 cells expressing BCR-ABL T315I mutation (IC 50; 11 nM). In PACE (Ponatinib Ph+ ALL and CML Evaluation) clinical trial indicated that 57% had CCyR (complete cytogenetical response), and 47% had MMR (major molecular response) in CML-chronic phase with T315I mutation. Ponatinib has substantial activity in heavily pretreated patients and those with refractory T315I mutation. However, approximately one third of ponatinib-treated patients had moderate to severe thrombocytopenia (J.E. Cortes et al. 2011 ASH Annual Meeting). The mechanism of ponatinib-induced thrombocytopenia remains unknown. In this study, we evaluated the effects of ponatinib on megakaryocytic progenitor cells, megakaryocytopoiesis, megakaryocyte and platelet production in mice. Method: All animal procedures were approved by the Institutional Animal Care and Use Committee of Iwate Medical University. Male ddY mice at 8 weeks of age were used in all experiments. We studied in vitro culture of megakaryocytic colonies (CFU-Meg), megakaryocyte ploidy analyses in vitro culture and proplatelet formation (PPF) assay in vitro. Murine bone marrow cells were cultured in methylcellulose with mIL-3, mIL-6 and TPO at 37°C in 5% CO2 and 20% O2 for 7 days in the presence of ponatinib (0.01, 0.1, 1, 10, 100 nM). PPF: Murine megakarocytes were partially purified from bone marrow cells using BSA gradient. They were plated in 96 micro-well culture plates (300 megakaryocyte/well) and cultured in IMDM, supplemented with 1% ITS-G (serum-free medium) in the presence of ponatinib (0.01, 0.1, 1, 10, 100 nM), at 37°C in 5% CO2and 20% O2. After 24 hr incubation, the megakaryocytes with proplatelets in each well were counted. Activated Rho and activated Rac in murine platelets were measured by the Western blot using Rhotekin-binding domain (RBD) beads and PAK-PBD Affinity beads respectively. The phosphorylation of Lyn (Src family kinase) in murine platelets was also evaluated by the Western blots. Results: CFU-Megs did not decrease significantly at 0.01, 0.1, 1, 10 and 100 nM (31.8 +/− 1.4 to 42.3 +/− 2.4 cells) and decreased significantly (17.0 +/− 1.6 cells p<0.01) at 1000 nM of ponatinib. PPF were decreased significantly at 0.1, 1, 10, 100 nM ponatinib (0 nM: 26.4 ± 0.8 %, 0.1 nM: 19.2 ± 1.7% p<0.05, 1 nM: 19.4 ± 2.1 % p<0.05, 10 nM: 17.9 ± 1.1% p<0.01, 100 nM: 12.5 ± 1.1 % p<0.001, 1000 nM: 11.6 ± 0.9 % p<0.001). The decreases in PPF were cancelled significantly by the addition of Y27632, Rho-associate kinase ROCK inhibitor (ponatinib 10 nM; 17.9 ± 1.1%, ponatinib 10 nM + Y27632 10μM; 29.8 ± 1.7% p<0.001, ponatinib 100 nM; 12.5 ± 1.1%, ponatinib 100 nM + Y27632 10μM; 19.3 ± 1.4% p<0.001). There was no difference in DNA ploidy of cultured megakaryocytes in the presence of ponatinib (0.01 to 100 nM). Next we have tried to understand the precise role of Rho/Rock and Rac pathways in platelets. Our data showed that Rho was upregulated and that activated Rac was downregulated at 50 nM of ponatinib. Ponatinib reduced the levels of phosphorylated Lyn (p-Lyn). Discussion and Conclusion: The Rho/ROCK pathway was reported to be negative regulators (Blood 2007 109; 4229) and positive regulators (Blood 2007 110; 3637) in PPF. Ponatinib induced thrombocytopenia might not be due to the inhibition of megakaryocyte colony formations but the inhibition of PPF of megakaryocytes via pathways including Rho/Rock and Rac. Disclosures: No relevant conflicts of interest to declare.

Fresh Bone Marrow and Periosteum Transplantation for Cartilage Defects of the Knee

2006 ◽  
Vol 38 (1) ◽  
pp. 318-319 ◽  
Author(s):  
K. Slynarski ◽  
J. Deszczynski ◽  
J. Karpinski
Keyword(s):  
Bone Marrow ◽  
Cartilage Defects ◽  
Fresh Bone

scholarly journals Expansion of Human Mesenchymal Stromal Cells from Fresh Bone Marrow in a 3D Scaffold-Based System under Direct Perfusion

PLoS ONE ◽  
2014 ◽  
Vol 9 (7) ◽  
pp. e102359 ◽  
Author(s):  
Adam Papadimitropoulos ◽  
Elia Piccinini ◽  
Sophie Brachat ◽  
Alessandra Braccini ◽  
David Wendt ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
3D Scaffold ◽  
Human Mesenchymal Stromal Cells ◽  
Fresh Bone

scholarly journals Mesenchymal content of fresh bone marrow: a proposed quality control method for cell therapy

2007 ◽  
Vol 139 (2) ◽  
pp. 312-320 ◽  
Author(s):  
Richard Veyrat-Masson ◽  
Nathalie Boiret-Dupré ◽  
Chantal Rapatel ◽  
Stéphane Descamps ◽  
Laurent Guillouard ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Quality Control ◽  
Cell Therapy ◽  
Control Method ◽  
Quality Control Method ◽  
Fresh Bone
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