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Neuroscience
Aislamiento de los capilares cerebrales del cerebro humano fresco tejido
Aislamiento de los capilares cerebrales del cerebro humano fresco tejido
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Isolation of Cerebral Capillaries from Fresh Human Brain Tissue

Aislamiento de los capilares cerebrales del cerebro humano fresco tejido

Full Text
13,367 Views
06:35 min
September 12, 2018

DOI: 10.3791/57346-v

Anika M.S. Hartz1, Julia A. Schulz2, Brent S. Sokola2, Stephanie E. Edelmann1, Andrew N. Shen1, Ralf G. Rempe2, Yu Zhong1, Nader El Seblani3, Bjoern Bauer2

1Sanders-Brown Center on Aging, Department of Pharmacology and Nutritional Sciences,University of Kentucky, 2Department of Pharmaceutical Sciences, College of Pharmacy,University of Kentucky, 3Department of Neuroscience,University of Kentucky

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study introduces a preclinical model using isolated brain capillaries from human brain tissue to investigate blood-brain barrier (BBB) function under both normal and pathological conditions. An optimized protocol for isolating these capillaries is detailed, enabling insights into BBB dysfunction and related signaling pathways.

Key Study Components

Area of Science

  • Neuroscience
  • Blood-Brain Barrier Research
  • Cellular Biology

Background

  • Blood-brain barrier integrity is critical for central nervous system function.
  • Understanding BBB dysfunction is pivotal for therapeutic development.
  • Existing models often lack the physiological relevance of human tissue.
  • This protocol leverages fresh human brain tissue for precise research applications.

Purpose of Study

  • To provide a reliable method for isolating human brain capillaries.
  • To study intact, physiologically active human capillaries.
  • To facilitate research into signaling pathways involved in BBB function.

Methods Used

  • The model involves isolating capillaries from fresh human brain tissue using a stepwise protocol.
  • Capillaries are separated from other brain debris using density gradient centrifugation.
  • Microscopic examination helps assess capillary purity post-isolation.
  • Capillaries can be stored for future experiments or processed immediately.
  • Safety measures for handling human tissue are emphasized due to infection risks.

Main Results

  • The method successfully isolates pure populations of human brain capillaries.
  • Insights on BBB modeling can further research into pharmacological effects and signaling pathways.
  • Isolation quality directly impacts the utility of capillaries in experimentation.
  • Potential applications include molecular, proteomic, and cellomic studies.

Conclusions

  • This study demonstrates a practical method for studying blood-brain barrier functionality in human tissue.
  • The approach enhances research capacity in understanding BBB-related mechanisms in health and disease.
  • Findings support advanced exploration of therapeutic avenues targeting barrier dysfunction.

Frequently Asked Questions

What are the advantages of using human brain capillaries?
Human brain capillaries provide a more relevant model for studying blood-brain barrier physiology and pathology compared to other species or in vitro systems.
How is the isolation protocol implemented?
The protocol involves careful dissection, density gradient centrifugation, and filtration to ensure high purity and yield of brain capillaries from tissue samples.
What types of experiments can isolated capillaries be used for?
Isolated capillaries can be used in pharmacological assays, cellular signaling studies, and various molecular analyses to investigate blood-brain barrier mechanisms.
What are key considerations during the isolation process?
Care must be taken to avoid capillary loss during handling, and the proper density of Ficoll is crucial for effective separation.
What safety precautions are necessary when working with human tissue?
Laboratory safety protocols include wearing protective gear, handling within a BSL-2 environment, and following guidelines for potential pathogen exposure.

Tubos capilares del cerebro aislado de tejido de cerebro humano pueden utilizarse como un modelo preclínico para estudiar la función de barrera bajo condiciones fisiológicas y fisiopatológicas. Aquí, presentamos un protocolo optimizado para aislar los capilares del cerebro del tejido de cerebro humano fresco.

Este método puede ayudar a responder preguntas clave en el campo de la investigación de la barrera hematoencefálica, como la disfunción de la barrera hematoencefálica, la fuga de la barrera y las vías de señalización específicas de la barrera hematoencefálica humana. La principal ventaja de esta técnica es que permite el estudio de capilares cerebrales humanos intactos y fisiológicamente activos que se han aislado de tejido cerebral humano fresco. Para comenzar este procedimiento, documente el peso del tejido cerebral humano, que debe ser de unos 10 gramos.

Bajo el microscopio, retire cuidadosamente todas las meninges con pinzas y use un bisturí para cortar la materia blanca. A continuación, corta con cuidado el tejido cerebral y pícalo con un bisturí durante unos cinco minutos. Transfiera el tejido cerebral al triturador de pañuelos y agregue 30 mililitros de tampón de aislamiento.

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