Activated macrophages as a feeder layer for growth of resident cardiac progenitor cells

Sepulveda, Diana E.; Meckert, Patricia Cabeza; Locatelli, Paola; Olea, Fernanda Daniela; Pérez, Néstor Gustavo; Pinilla, Oscar Andrés; Díaz, Romina Gisel; Crottogini, Alberto J.; Laguens, Rubén M.

doi:10.1007/s10616-014-9814-0

Cited by 3 publications

(3 citation statements)

References 24 publications

(32 reference statements)

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“…However, we found that the fibrotic myocardium is still detected in the splenectomy/telmisartan-treated animals. Therefore, we cannot exclude the possibility that progenitor macrophages released from the bone marrow or resident macrophages in the heart may also participate in the Ang II-stimulated cardiac fibrosis, 23 , 24 which may not be altered by splenectomy/telmisartan. Furthermore, using flow cytometry may help to quantitatively distinguish subtypes of monocytes in the spleen and demonstrate the surface marker expression of monocytes after the AT1 receptor stimulation.…”

Section: Discussionmentioning

confidence: 99%

Recruitment of macrophages from the spleen contributes to myocardial fibrosis and hypertension induced by angiotensin II

Wang

Erskine

Zhang

et al. 2017

J Renin Angiotensin Aldosterone Syst

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Introduction:The purpose of this study was to determine whether macrophages migrated from the spleen are associated with angiotensin II-induced cardiac fibrosis and hypertension.Methods:Sprague-Dawley rats were subjected to angiotensin II infusion in vehicle (500 ng/kg/min) for up to four weeks. In splenectomy, the spleen was removed before angiotensin II infusion. In the angiotensin II AT1 receptor blockade, telmisartan was administered by gastric gavage (10 mg/kg/day) during angiotensin II infusion. The heart and aorta were isolated for Western blot analysis and immunohistochemistry.Results:Angiotensin II infusion caused a significant reduction in the number of monocytes in the spleen through the AT1 receptor-activated monocyte chemoattractant protein-1. Comparison of angiotensin II infusion, splenectomy and telmisartan comparatively reduced the recruitment of macrophages into the heart. Associated with this change, transforming growth factor β1 expression and myofibroblast proliferation were inhibited, and Smad2/3 and collagen I/III were downregulated. Furthermore, interstitial/perivascular fibrosis was attenuated. These modifications occurred in coincidence with reduced blood pressure. At week 4, invasion of macrophages and myofibroblasts in the thoracic aorta was attenuated and expression of endothelial nitric oxide synthase was upregulated, along with a reduction in aortic fibrosis.Conclusions:These results suggest that macrophages when recruited into the heart and aorta from the spleen potentially contribute to angiotensin II-induced cardiac fibrosis and hypertension.

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Section: Discussionmentioning

confidence: 99%

Recruitment of macrophages from the spleen contributes to myocardial fibrosis and hypertension induced by angiotensin II

Wang

Erskine

Zhang

et al. 2017

J Renin Angiotensin Aldosterone Syst

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“…Preparation of feeder cells. Peritoneal macrophages (PM) were used as feeder cells, after being obtained according to a previously published protocol (13). Briefly, Kunming mice were intraperitoneally injected with 5 mL of thioglycolate.…”

Section: Methodsmentioning

confidence: 99%

Cloning and identification of PK15 cells for enhanced replication of classical swine fever virus

Yin

et al. 2020

Journal of Veterinary Research

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IntroductionClassical swine fever virus (CSFV) causes an economically important and highly contagious disease of pigs, leading to economic losses around the world. Attenuated live vaccines with CSFV antigens have played an important role in the prevention and control of the disease. Porcine kidney 15 (PK15) cells have been widely used for the propagation of CSFV, but this cell line is not efficient or homogeneously susceptible to viral infection.Material and MethodsTo achieve a homogeneous PK15 cell line which enabled high titre replication of CSFV, we used the limiting dilution cell cloning method.ResultsWe developed two cell clones, PK15-1A6 and PK15-3B1, which respectively have high- and low-permissive phenotypes to CSFV infection. The PK15-1A6, PK15-3B1, and PK15 parent cells showed different characteristics in cell proliferation rate, susceptibility to CSFV infection, and CSFV production. The mean virus titres per millilitre reflected by TCID50 values in PK15-1A6, PK15-3B1, and PK15 parent cells were 106.85, 103.63, and 104.74, respectively.ConclusionThe PK15-1A6 cell clone is more permissive to CSFV infection than the PK15 parent cells. The screened high-permissive cells will be useful for CSFV propagation and vaccine development in vitro, and facilitate research on the pathogenicity of CSFV.

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“…Seven days previous to diaphragm extraction, 20 cm 3 of blood were obtained from each sheep's antecubital vein in order to construct a feeder layer of autologous macrophages, as previously reported (Sepúlveda et al 2016). This feeder layer was employed only for the primary culture of DMs but was not needed after the first passage.…”

Section: Obtention Of a Feeder Layer Of Autologous Macrophagesmentioning

confidence: 99%

Aligned ovine diaphragmatic myoblasts overexpressing human connexin-43 seeded on poly (l-lactic acid) scaffolds for potential use in cardiac regeneration

et al. 2017

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Diaphragmatic myoblasts (DMs) are precursors of type-1 muscle cells displaying high exhaustion threshold on account that they contract and relax 20 times/min over a lifespan, making them potentially useful in cardiac regeneration strategies. Besides, it has been shown that biomaterials for stem cell delivery improve cell retention and viability in the target organ. In the present study, we aimed at developing a novel approach based on the use of poly (L-lactic acid) (PLLA) scaffolds seeded with DMs overexpressing connexin-43 (cx43), a gap junction protein that promotes inter-cell connectivity. DMs isolated from ovine diaphragm biopsies were characterized by immunohistochemistry and ability to differentiate into myotubes (MTs) and transduced with a lentiviral vector encoding cx43. After confirming cx43 expression (RT-qPCR and Western blot) and its effect on inter-cell connectivity (fluorescence recovery after photobleaching), DMs were grown on fiber-aligned or random PLLA scaffolds. DMs were successfully isolated and characterized. Cx43 mRNA and protein were overexpressed and favored inter-cell connectivity. Alignment of the scaffold fibers not only aligned but also elongated the cells, increasing the contact surface between them. This novel approach is feasible and combines the advantages of bioresorbable scaffolds as delivery method and a cell type that on account of its features may be suitable for cardiac regeneration. Future studies on animal models of myocardial infarction are needed to establish its usefulness on scar reduction and cardiac function.

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Activated macrophages as a feeder layer for growth of resident cardiac progenitor cells

Cited by 3 publications

References 24 publications

Recruitment of macrophages from the spleen contributes to myocardial fibrosis and hypertension induced by angiotensin II

Recruitment of macrophages from the spleen contributes to myocardial fibrosis and hypertension induced by angiotensin II

Cloning and identification of PK15 cells for enhanced replication of classical swine fever virus

Aligned ovine diaphragmatic myoblasts overexpressing human connexin-43 seeded on poly (l-lactic acid) scaffolds for potential use in cardiac regeneration

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