Current Activities

• Blog Stats

  • 3,509 hits

• Poster Text as presented at the 2013 World Congress of Neurology

  A BLOOD BRAIN BARRIER BIOREPOSITORY FOR DISEASE-SPECIFIC HUMAN BRAIN MICROVASCULAR ENDOTHELIAL CELLS: A NEW TOOL FOR NEUROLOGIC AND PSYCHIATRIC RESEARCH

  Thomas R. Roesel MD, PhD

  Cervoe.org

  OBJECTIVE
  The objective is two-fold: 1) to confirm the need for a biorepository for disease-specific human brain microvessel endothelial cells (HBMECs), and 2) to present a schematic for establishing the first blood brain barrier (BBB) bank to make available disease-specific HBMECs for clinically relevant translational medical research.

  BACKGROUND
  BBB dysfunction has been implicated in many illnesses of the central nervous system such as stroke (1,2), traumatic brain and spine injury (2,3,4), Parkinson's disease (5), Alzheimer's disease (6), schizophrenia (2,7,8), multiple sclerosis (9), as well as more uncommon conditions such as chronic Lyme disease (10,11), chronic fatigue syndrome (12), Gulf War-related illness (13), and human immune deficiency virus (HIV) associated dementia (HAD) (6,14). Investigations on BBB pathophysiology are restricted by the inability to construct in vitro BBB culture experiments using disease-specific HBMECs due to the lack of their on demand availability, despite the routine availability of normal, non-disease related HBMECs.

  METHODS
  Non-oncological brain tissue banks throughout the world were identified through literature and website searches. These repositories were evaluated to determine whether their services included the isolation of disease specific HBMECs for dissemination to researchers not affiliated with the HBMEC procurement process. This information was obtained by scrutiny of the identified biorepository website and/or by direct email query to that repository. Furthermore, the scientific literature was reviewed for HBMEC isolation and cryopreservation techniques, as routinely utilized by HBMEC providers for the procurement of normal, nonspecific HBMECs, to develop a protocol for the banking of disease-specific HBMECs.

  RESULTS
  Seventy-nine non-oncological brain banks or their umbrella networks in 16 different countries were identified by literature and website searches. In addition, 8 commercial vendors for HBMECs were similarly identified. Neither the commercial or non-commercial brain tissue banks offered disease-specific HBMECs for laboratory investigations. However, 11 non-commercial banks could provide disease-specific HBMECs with advanced notification, should a disease appropriate donor become available.

  DISCUSSION
  With the lack of availability of a biorepository committed to supplying disease-specific HBMECs on demand, a construct for such a repository was developed. The proposed biorepository is based on current principles that govern established tissue bank repositories as followed by commercial and non-commercial vendors which provide HBMECs that lack disease specificity (15). Biorepository functions are divided into administrative, laboratory, and cryopreservation storage areas. Personnel can have overlapping duties in each of these areas, but physical barriers between the designated areas should exist to avoid sample contamination. Administrators help to recruit volunteer donors, obtain pre-endowment consent, and acquire medical history before and at the time of death. Post-death consent from the family is required and donor encryption principles are followed. Administrators also disseminate to the investigator, who request samples, the medical history related to the disease-specific HBMECs under policies that assure donor privacy. Tissue acquisition and HBMEC isolation follow established procedures for all volunteer brain endowments (16,17). Tissue and cell handling, assessment and documentation of the “state of the brain,” including photography, and subsequent dissection of the cerebral brain microvessels are performed by laboratory personnel. The microvessels from disease affected areas of the brain as well as non-disease affected control areas, when necessary, are processed so that HBMECs are isolated. These disease- specific HBMECs are then placed into appropriately labeled vials and cryopreserved for storage by standard procedures until dissemination to the requesting investigators (16,17). Donor confidentiality is maintained at all times.

  CONCLUSIONS
  Establishment of a biorepository for cryopreserved disease-specific HBMECs is feasible, and can provide needed cells for BBB-related translational medicine studies in neurology and psychiatry. There are currently no such banks dedicated to providing disease-specific HBMECs. Using such cells to increase pathophysiologic knowledge and to discover interventions to increase BBB resiliency for specific diseases can be explored with on-demand disease-specific HBMECs. Efforts are underway to establish the first such biorepository.

  REFERENCES
  1. Sandoval KE, Witt KA. Blood-brain barrier tight junction permeability and ischemic stroke. Neurobiol Dis 2008; 32:200-209.
  2. Schoknecht K, Shalev H. Blood-brain barrier dysfunction in brain diseases: Clinical experience. Epilesia 2012; 53 (Suppl. 6):7-13.
  3. Chodobski A, Zink BJ, Szmydynger-Chodobska. Blood brain pathophysiology in traumatic brain injury. Transl Stroke Res 2011; 2(4): 492-516.
  4. Marchi N, Bazarian JJ, Puvenna V, et al. Consequences of repeated blood-brain barrier disruption in football players. PLoS ONE 2013; 8(3): e56805
  5. Kortekaas R, Leenders KL, van Oostrom JC, et al. Blood-brain barrier dysfunction in parkinsonian midbrain in vivo. Ann Neurol. 2005; 57:176-179.
  6. Idaecola C. The overlap between neurodegenerative and vascular factors in the pathogenesis of dementia. Acta Neuropathol 2010; 120(3):287-296.
  7. Uranova NA, Zimina IS, Vikhreva OV, et al. Ultrastructural damage of capillaries in the neocortex in schizophrenia. The World Journal of Biol Psych 2010; 11:567-578.
  8. Harris LW, Wayland M, Lan M, et al. The cerebral microvasculature in schizophrenia: A laser capture microdissection study. PLoS ONE 2008; 3(12): e3964.
  9. Hemmer B, Cepok S, Zhou D, Sommer N. Multiple Sclerosis—a coordinated immune attack across the blood brain barrier. Curr Neurovasc Res 2004; 1:141-150.
  10. Choi S, Stins MF, Kim KS, Kim J, et al. Borellia burgdorferi, host-derived proteases, and the blood-brain barrier. Infect Immun 2005; 73(2):1014-1022.
  11. Brendtson, K. Review of the evidence for immune evasion and persistent infections in Lyme disease. Int J of Gen Med 2013; 6:291-306.
  12. Bested AC, Saunders PR, Logan AC. Chronic fatigue syndrome: neurological findings may be related to blood-brain barrier permeability. Med Hypotheses 2001; 57(2):231-237.
  13. Abdel-Rahman A, Shetty AK, Abou-Donia MB. Disruption of the blood-brain barrier and neuronal cell death in cingulate cortex, dentate gyrus, thalamus, and hypothalamus in a rat model of Gulf-War syndrome. Neurobiol Dis 2002; 10(3):306-326.
  14. Chaudhuri A, Duan F, Morsey B, et al. HIV-1 activates proinflammatory and interferon-inducible genes in human brain microvascular endothelial cells: putative mechanisms of blood-brain barrier dysfunction. J Cerebral Blood Flow & Metab 2008; 28:697-711.
  15. Fehily D, Brubaker S, Kearney JN, Wolfinbarger L, (editors). Tissue and cell processing: an essential guide. Chichester, United Kingdom: John Wiley & Sons 2012.
  16. Biegel D, Spencer DD, Pachter JS. Isolation and culture of human brain microvessel cells for the study of blood-brain barrier properties in vitro. Brain Res 1995; 692(1-2):183-189.
  17. Dorovini-Zis K, Prameya R, Huynh H. Isolation and characterization of human brain endothelial cells. Methods in Mol Med 2003; 89:325-336.