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Secretomes of apoptotic mononuclear cells ameliorate neurological damage in rats with focal ischemia.

Altmann P, Mildner M, Haider T, Traxler D, Beer L, Ristl R, Golabi B, Gabriel C, Leutmezer F, Ankersmit HJ - F1000Res (2014)

Bottom Line: Administration of rat as well as human apoptotic MNC-secretomes significantly reduced ischemic lesion volumes by 36% and 37%, respectively.Co-incubation of human astrocytes, Schwann cells and neurons with hMNC (apo sec) resulted in activation of several signaling cascades associated with the regulation of cytoprotective gene products and enhanced neuronal sprouting in vitro.Analysis of neurotrophic factors in hMNC (apo sec) and rat plasma revealed high levels of brain derived neurotrophic factor (BDNF).

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Medical University of Vienna, Vienna, 1090, Austria ; Christian Doppler Laboratory for Cardiac and Thoracic Diagnosis and Regeneration, Vienna, 1090, Austria.

ABSTRACT
The pursuit of targeting multiple pathways in the ischemic cascade of cerebral stroke is a promising treatment option. We examined the regenerative potential of conditioned medium derived from rat and human apoptotic mononuclear cells (MNC), rMNC (apo sec) and hMNC (apo sec), in experimental stroke. We performed middle cerebral artery occlusion on Wistar rats and administered apoptotic MNC-secretomes intraperitoneally in two experimental settings. Ischemic lesion volumes were determined 48 hours after cerebral ischemia. Neurological evaluations were performed after 6, 24 and 48 hours. Immunoblots were conducted to analyze neuroprotective signal-transduction in human primary glia cells and neurons. Neuronal sprouting assays were performed and neurotrophic factors in both hMNC (apo sec) and rat plasma were quantified using ELISA. Administration of rat as well as human apoptotic MNC-secretomes significantly reduced ischemic lesion volumes by 36% and 37%, respectively. Neurological examinations revealed improvement after stroke in both treatment groups. Co-incubation of human astrocytes, Schwann cells and neurons with hMNC (apo sec) resulted in activation of several signaling cascades associated with the regulation of cytoprotective gene products and enhanced neuronal sprouting in vitro. Analysis of neurotrophic factors in hMNC (apo sec) and rat plasma revealed high levels of brain derived neurotrophic factor (BDNF). Our data indicate that apoptotic MNC-secretomes elicit neuroprotective effects on rats that have undergone ischemic stroke.

No MeSH data available.


Related in: MedlinePlus

Experimental study setting.For setting 1, rMNCapo sec (apoptotic MNC-secretomes from rats) were injected 40 minutes after MCAO (blue arrow). In setting 2, hMNCapo sec (apoptotic MNC-secretomes from humans) were administered twice at 40 minutes (0.7 hours) and 24 hours after MCAO (red arrows). In both settings, neurological evaluations were performed at 0 hours (before surgery) as well as 6, 24, and 48 hours after surgery (boxes). Both treatment and control animals were euthanized 48 hours after surgery and brain slices were treated with TTC (2,3,5-triphenyltetrazolium chloride) to stain ischemic areas in the brain.
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f1: Experimental study setting.For setting 1, rMNCapo sec (apoptotic MNC-secretomes from rats) were injected 40 minutes after MCAO (blue arrow). In setting 2, hMNCapo sec (apoptotic MNC-secretomes from humans) were administered twice at 40 minutes (0.7 hours) and 24 hours after MCAO (red arrows). In both settings, neurological evaluations were performed at 0 hours (before surgery) as well as 6, 24, and 48 hours after surgery (boxes). Both treatment and control animals were euthanized 48 hours after surgery and brain slices were treated with TTC (2,3,5-triphenyltetrazolium chloride) to stain ischemic areas in the brain.

Mentions: In the first experimental setting, lyophilized rMNCapo sec (produced from 12.5×106 apoptotic rat MNCs) or control medium was resuspended in 0.3 mL saline (Fresenius Kabi, Vienna, Austria) in the laboratory prior to surgery. In order to investigate the potency of apoptotic MNC-secretomes, both the treatment group and the control group (n=29) randomly received 0.3 mL rMNCapo sec or control medium intraperitoneally forty minutes after surgery (Figure 1, blue arrow). In the second setting, lyophilized hMNCapo sec (produced from 12.5×106 apoptotic human MNCs) or lyophilized control medium were each resuspended in the laboratory in 0.3 mL saline prior to surgery. In order to investigate whether a higher dosage and time interval would provide additional benefits, animals from experimental setting 2 (n=29) received two intraperitoneal doses 40 minutes and 24 hours after MCAO induction (Figure 1, red arrows). The rationale for this two-step-approach is given in the discussion. In addition, all animals were given a subcutaneous injection of 3.5 mL/kg saline after surgery and put under a heating lamp until they woke up.


Secretomes of apoptotic mononuclear cells ameliorate neurological damage in rats with focal ischemia.

Altmann P, Mildner M, Haider T, Traxler D, Beer L, Ristl R, Golabi B, Gabriel C, Leutmezer F, Ankersmit HJ - F1000Res (2014)

Experimental study setting.For setting 1, rMNCapo sec (apoptotic MNC-secretomes from rats) were injected 40 minutes after MCAO (blue arrow). In setting 2, hMNCapo sec (apoptotic MNC-secretomes from humans) were administered twice at 40 minutes (0.7 hours) and 24 hours after MCAO (red arrows). In both settings, neurological evaluations were performed at 0 hours (before surgery) as well as 6, 24, and 48 hours after surgery (boxes). Both treatment and control animals were euthanized 48 hours after surgery and brain slices were treated with TTC (2,3,5-triphenyltetrazolium chloride) to stain ischemic areas in the brain.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215751&req=5

f1: Experimental study setting.For setting 1, rMNCapo sec (apoptotic MNC-secretomes from rats) were injected 40 minutes after MCAO (blue arrow). In setting 2, hMNCapo sec (apoptotic MNC-secretomes from humans) were administered twice at 40 minutes (0.7 hours) and 24 hours after MCAO (red arrows). In both settings, neurological evaluations were performed at 0 hours (before surgery) as well as 6, 24, and 48 hours after surgery (boxes). Both treatment and control animals were euthanized 48 hours after surgery and brain slices were treated with TTC (2,3,5-triphenyltetrazolium chloride) to stain ischemic areas in the brain.
Mentions: In the first experimental setting, lyophilized rMNCapo sec (produced from 12.5×106 apoptotic rat MNCs) or control medium was resuspended in 0.3 mL saline (Fresenius Kabi, Vienna, Austria) in the laboratory prior to surgery. In order to investigate the potency of apoptotic MNC-secretomes, both the treatment group and the control group (n=29) randomly received 0.3 mL rMNCapo sec or control medium intraperitoneally forty minutes after surgery (Figure 1, blue arrow). In the second setting, lyophilized hMNCapo sec (produced from 12.5×106 apoptotic human MNCs) or lyophilized control medium were each resuspended in the laboratory in 0.3 mL saline prior to surgery. In order to investigate whether a higher dosage and time interval would provide additional benefits, animals from experimental setting 2 (n=29) received two intraperitoneal doses 40 minutes and 24 hours after MCAO induction (Figure 1, red arrows). The rationale for this two-step-approach is given in the discussion. In addition, all animals were given a subcutaneous injection of 3.5 mL/kg saline after surgery and put under a heating lamp until they woke up.

Bottom Line: Administration of rat as well as human apoptotic MNC-secretomes significantly reduced ischemic lesion volumes by 36% and 37%, respectively.Co-incubation of human astrocytes, Schwann cells and neurons with hMNC (apo sec) resulted in activation of several signaling cascades associated with the regulation of cytoprotective gene products and enhanced neuronal sprouting in vitro.Analysis of neurotrophic factors in hMNC (apo sec) and rat plasma revealed high levels of brain derived neurotrophic factor (BDNF).

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Medical University of Vienna, Vienna, 1090, Austria ; Christian Doppler Laboratory for Cardiac and Thoracic Diagnosis and Regeneration, Vienna, 1090, Austria.

ABSTRACT
The pursuit of targeting multiple pathways in the ischemic cascade of cerebral stroke is a promising treatment option. We examined the regenerative potential of conditioned medium derived from rat and human apoptotic mononuclear cells (MNC), rMNC (apo sec) and hMNC (apo sec), in experimental stroke. We performed middle cerebral artery occlusion on Wistar rats and administered apoptotic MNC-secretomes intraperitoneally in two experimental settings. Ischemic lesion volumes were determined 48 hours after cerebral ischemia. Neurological evaluations were performed after 6, 24 and 48 hours. Immunoblots were conducted to analyze neuroprotective signal-transduction in human primary glia cells and neurons. Neuronal sprouting assays were performed and neurotrophic factors in both hMNC (apo sec) and rat plasma were quantified using ELISA. Administration of rat as well as human apoptotic MNC-secretomes significantly reduced ischemic lesion volumes by 36% and 37%, respectively. Neurological examinations revealed improvement after stroke in both treatment groups. Co-incubation of human astrocytes, Schwann cells and neurons with hMNC (apo sec) resulted in activation of several signaling cascades associated with the regulation of cytoprotective gene products and enhanced neuronal sprouting in vitro. Analysis of neurotrophic factors in hMNC (apo sec) and rat plasma revealed high levels of brain derived neurotrophic factor (BDNF). Our data indicate that apoptotic MNC-secretomes elicit neuroprotective effects on rats that have undergone ischemic stroke.

No MeSH data available.


Related in: MedlinePlus