Bone Marrow-Derived Mesenchymal Stem Cells: Isolation, Expansion, Characterization, Viral Transduction, and Production of Conditioned Medium

2009 ◽  
pp. 281-294 ◽  
Author(s):  
Massimiliano Gnecchi ◽  
Luis G. Melo
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Viral Transduction

scholarly journals Correction: Conditioned Medium from Hypoxic Bone Marrow-Derived Mesenchymal Stem Cells Enhances Wound Healing in Mice

PLoS ONE ◽  
2015 ◽  
Vol 10 (12) ◽  
pp. e0145565 ◽  
Author(s):  
Lei Chen ◽  
Yingbin Xu ◽  
Jingling Zhao ◽  
Zhaoqiang Zhang ◽  
Ronghua Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium

scholarly journals Patient Heterogeneity in Acute Myeloid Leukemia: Leukemic Cell Communication by Release of Soluble Mediators and Its Effects on Mesenchymal Stem Cells

Diseases ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 74
Author(s):  
Elise Aasebø ◽  
Annette K. Brenner ◽  
Maria Hernandez-Valladares ◽  
Even Birkeland ◽  
Olav Mjaavatten ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Extracellular Matrix ◽  
Acute Myeloid Leukemia ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Untreated Control ◽  
Myeloid Leukemia ◽  
Control Mscs ◽  
Acute Myeloid

Acute myeloid leukemia (AML) is an aggressive bone marrow malignancy, and non-leukemic stromal cells (including mesenchymal stem cells, MSCs) are involved in leukemogenesis and show AML-supporting effects. We investigated how constitutive extracellular mediator release by primary human AML cells alters proteomic profiles of normal bone marrow MSCs. An average of 6814 proteins (range 6493−6918 proteins) were quantified for 41 MSC cultures supplemented with AML-cell conditioned medium, whereas an average of 6715 proteins (range 6703−6722) were quantified for untreated control MSCs. The AML effect on global MSC proteomic profiles varied between patients. Hierarchical clustering analysis identified 10 patients (5/10 secondary AML) showing more extensive AML-effects on the MSC proteome, whereas the other 31 patients clustered together with the untreated control MSCs and showed less extensive AML-induced effects. These two patient subsets differed especially with regard to MSC levels of extracellular matrix and mitochondrial/metabolic regulatory proteins. Less than 10% of MSC proteins were significantly altered by the exposure to AML-conditioned media; 301 proteins could only be quantified after exposure to conditioned medium and 201 additional proteins were significantly altered compared with the levels in control samples (153 increased, 48 decreased). The AML-modulated MSC proteins formed several interacting networks mainly reflecting intracellular organellar structure/trafficking but also extracellular matrix/cytokine signaling, and a single small network reflecting altered DNA replication. Our results suggest that targeting of intracellular trafficking and/or intercellular communication is a possible therapeutic strategy in AML.

scholarly journals Conditioned Medium from Human Tonsil-Derived Mesenchymal Stem Cells Enhances Bone Marrow Engraftment via Endothelial Cell Restoration by Pleiotrophin

Cells ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 221
Author(s):  
Yu-Hee Kim ◽  
Kyung-Ah Cho ◽  
Hyun-Ji Lee ◽  
Minhwa Park ◽  
Sang-Jin Shin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Histological Analysis ◽  
Conditioning Regimen ◽  
Mouse Bone Marrow ◽  
Human Tonsil ◽  
Hematopoietic Stem ◽  
Bone Marrow Engraftment

Cotransplantation of mesenchymal stem cells (MSCs) with hematopoietic stem cells (HSCs) has been widely reported to promote HSC engraftment and enhance marrow stromal regeneration. The present study aimed to define whether MSC conditioned medium could recapitulate the effects of MSC cotransplantation. Mouse bone marrow (BM) was partially ablated by the administration of a busulfan and cyclophosphamide (Bu–Cy)-conditioning regimen in BALB/c recipient mice. BM cells (BMCs) isolated from C57BL/6 mice were transplanted via tail vein with or without tonsil-derived MSC conditioned medium (T-MSC CM). Histological analysis of femurs showed increased BM cellularity when T-MSC CM or recombinant human pleiotrophin (rhPTN), a cytokine readily secreted from T-MSCs with a function in hematopoiesis, was injected with BMCs. Microstructural impairment in mesenteric and BM arteriole endothelial cells (ECs) were observed after treatment with Bu–Cy-conditioning regimen; however, T-MSC CM or rhPTN treatment restored the defects. These effects by T-MSC CM were disrupted in the presence of an anti-PTN antibody, indicating that PTN is a key mediator of EC restoration and enhanced BM engraftment. In conclusion, T-MSC CM administration enhances BM engraftment, in part by restoring vasculature via PTN production. These findings highlight the potential therapeutic relevance of T-MSC CM for increasing HSC transplantation efficacy.

Investigation of the Influence of the Conditioning Medium Factors Obtained During the Cultivation of Bone Marrow Mesenchymal Stem Cells on the Course of Severe Local Radiation Injuries of Skin in Rats

2018 ◽  
Vol 63 (1) ◽  
pp. 35-43 ◽  
Author(s):  
А. Темнов ◽  
A. Temnov ◽  
Т. Астрелина ◽  
T. Astrelina ◽  
К. Рогов ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Skin Lesion ◽  
Conditioned Medium ◽  
Radiation Injuries ◽  
Lesion Area ◽  
Paracrine Factors ◽  
Local Radiation ◽  
Radiation Ulcers

Purpose: Study of the effect of paracrine factors, produced by MMSC of bone marrow during the cultivation, on the severity of local radiation injuries in the conditions of application in the early periods after irradiation. Material and methods: Experiments were performed on rats of the breed Wistar weighing 280 g. Rats were exposed locally in iliolumbar region of the back using X-ray machine LNC-268 (RAP 100-10) at a dose of 110 Gy (30 kV tube voltage, current 6.1 mA, filter Al 0.1 mm thick), dose rate is 21.4 Gy/min. Area of the irradiation field was 8.2–8.5 cm2. The conditioned medium obtained by culturing MMSC of rats’ bone marrow was administered in dose 1.0 ml (total protein 8 mg/ml) at 1, 3, 6, 8 and 10 days after irradiation. The severity of radiation damage to the skin and the effects of therapy were evaluated in dynamics by clinical manifestations, using planimetry and histological methods. Results: It was shown that in control animals and in rats, with the introduction of the conditioned medium, the values of the skin lesion area in the period up to the 29th day after irradiation practically did not differ, gradually decreasing in control animals from 5.9 ± 0.6 cm2 to 2.2 ± 0.3 cm2 at the 15th and 29th days after irradiation, respectively. Then, in the control group, the lesion area ranged from 1.4 ± 0.6 cm2 on the 50th day to 1.9 ± 0.8 cm2 on the 71st day. In the experimental group of animals, with the introduction of factors of the conditioning medium, a decrease in the area of the lesion and a stable dynamics of healing of radiation ulcers, beginning from the 36th day, there was a gradual decrease in the area of the lesion, which reached 0.2 ± 0.1 cm2 by the 71st day after irradiation. On the 64–71th day after irradiation, the difference between the areas of skin lesion in the experimental and control groups was statistically significant, p <0.05. The histological analysis showed that the use of paracrine factors obtained from MMSC in the process of cultivation significantly reduces the severity of the inflammatory reaction and accelerates the regeneration processes. Conclusion: Thus, the introduction of conditioned medium factors obtained during the cultivation of mesenchymal stem cells of the bone marrow facilitates a more easy flow of the pathological process and the healing of radiation ulcers after local radiation damage to the skin of rats. Apparently, the favorable effect of paracrine factors introduced in the early periods after irradiation, with severe local radiation injuries, is associated with their effect on pathological processes in the inflammatory-destructive stage.

scholarly journals Conditioned Medium from Hypoxic Bone Marrow-Derived Mesenchymal Stem Cells Enhances Wound Healing in Mice

PLoS ONE ◽  
2014 ◽  
Vol 9 (4) ◽  
pp. e96161 ◽  
Author(s):  
Lei Chen ◽  
Yingbin Xu ◽  
Jingling Zhao ◽  
Zhaoqiang Zhang ◽  
Ronghua Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium

scholarly journals Conditioned Medium of Bone Marrow Mesenchymal Stem Cells Relieves Acute Lung Injury in Mice by Regulating Epithelial Sodium Channels via miR-34c

2020 ◽  
Author(s):  
Zhiyu Zhou ◽  
Yong Cui ◽  
Yapeng Hou ◽  
Tong Yu ◽  
Yan Ding ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Acute Lung Injury ◽  
Lung Injury ◽  
Conditioned Medium ◽  
Luciferase Reporter ◽  
Rt Pcr ◽  
Wide Range ◽  
Gene Assay

Abstract Aims: One of the characteristics of acute lung injury (ALI) is severe pulmonary edema, which is closelyrelated to alveolar fluid clearance. Mesenchymal stem cells (MSCs) secrete a wide range of cytokines,growth factors and miRNAs through paracrine action to participate in the mechanism of pulmonaryinflammatory response, which increases the clearance of edema fluid, and promotes the repair process ofALI. However, the mechanism by which bone marrow derived MSCs-conditioned medium (BMSCs-CM)promotes edema clearance is unclear. Epithelial sodium channel (ENaC) is the rate-limiting step in thesodium-water transport and edema clearance in the alveolar cavity, and we aim to explore the role of ENaCin BMSCs-CM invloved edema clearance and whether it can alter the function of ENaC via miRNAs.Methods: CCK-8 cell proliferation assay was used to detect the effect of BMSCs-CM on the survival ofAT2 cells. Real-time PCR (RT-PCR) and Western blot were used to detect the expression of ENaC in AT2cells. The effects of exosomes/miR-34c on the transepithelial short-circuit current in the monolayer of H441cells were examined by the Ussing chamber setup. Dual luciferase reporter gene assay was used to detect thetarget gene of miR-34c.Results: BMSCs-CM can increase the viability of mouse AT2 cells. RT-PCR and Western blotting resultsshowed that BMSCs-CM significantly increased the expression of γ-ENaC subunit in mouse AT2 cells.Ussing chamber assay revealed that BMSCs-CM enhanced the amiloride-sensitive currents associated withENaC activity in intact H441 cell monolayers. In addition, we observed higher expression of miR-34c inmouse AT2 cells administrated with BMSCs-CM, and the overexpression or inhibition of miR-34c canregulate the expression of ENaC protein and alter the function of ENaC. Finally, we detected MARCKS maybe one of the target gene of miR-34c.Conclusions: Our results indicate that BMSCs-CM may improve LPS-induced ALI through miR-34ctargeting MARCKS and regulating ENaC indirectly, which further explores the benefit of paracrine effectsof BMSCs on edematous ALI.

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