Development, characterization and potential applications of a multicellular spheroidal human blood-brain barrier model integrating three conditionally immortalized cell lines
نویسندگان
چکیده
In vitro blood–brain barrier (BBB) models are essential research tools for use in developing brain-targeted drugs and understanding the physiological pathophysiological functions of BBB. To develop BBB with better functionalities, three-dimensional (3D) culture methods have gained significant attention as a promising approach. this study, we report on development human conditionally immortalized cell-based multicellular spheroidal (hiMCS-BBB) model. After being seeded into non-attachment wells, HASTR/ci35 (astrocytes) HBPC/ci37 cells (brain pericytes) self-assemble to form spheroid core that is then covered an outer monolayer HBMEC/ci18 microvascular endothelial cells). The results immunocytochemistry showed protein expression several cellular junction BBB-enriched transporter genes permeability assays hiMCS-BBB model exhibited against penetration dextran (5 70 kDa) rhodamine123 (a P-glycoprotein substrate) core. On other hand, facilitation 2-(N-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]amino)-2-deoxyglucose (2-NBDG; fluorescent glucose analog) uptake was observed Furthermore, tumor necrosis factor-alpha treatment elicited inflammatory response cells, thereby suggesting inflammation can be recapitulated summarize, developed possesses fundamental properties, which expected provide useful highly accessible experimental platform accelerating various studies.
منابع مشابه
Multicellular Self-Assembled Spheroidal Model of the Blood Brain Barrier
The blood brain barrier (BBB) has evolved unique characteristics such as dense coverage of the endothelial cells by pericytes and interactions with astrocytes through perivascular endfeet. We study BBB formation in a 3-dimensional multicellular spheroid system of human primary brain endothelial cells (hpBECs), primary pericytes (hpPs) and primary astrocytes (hpAs). We show for the first time th...
متن کاملConditionally immortalized cell lines as a new in vitro model for the study of barrier functions.
Conditionally immortalized brain and retinal capillary endothelial and choroid plexus epithelial cell lines were established from a transgenic rat (Tg rat) and mouse (Tg mouse) harboring the temperature-sensitive simian virus 40 (ts SV 40) large T-antigen. These cell lines exhibit temperature-sensitive cell growth due to the expression of ts SV 40 large T-antigen. Mouse brain (TM-BBB) and rat b...
متن کاملDevelopment of a conditionally immortalized human pancreatic β cell line.
Diabetic patients exhibit a reduction in β cells, which secrete insulin to help regulate glucose homeostasis; however, little is known about the factors that regulate proliferation of these cells in human pancreas. Access to primary human β cells is limited and a challenge for both functional studies and drug discovery progress. We previously reported the generation of a human β cell line (Endo...
متن کاملConditionally immortalized human podocyte cell lines established from urine.
Evidence suggests that loss of podocytes into urine contributes to development of glomerular diseases; shed podocytes are frequently viable and proliferate in culture conditions. To determine the phenotypic characteristics of viable urinary cells derived from human subjects, we established long-term urinary cell culture from two patients with focal segmental glomerulosclerosis and two healthy v...
متن کاملA retinoic acid-enhanced, multicellular human blood-brain barrier model derived from stem cell sources
Blood-brain barrier (BBB) models are often used to investigate BBB function and screen brain-penetrating therapeutics, but it has been difficult to construct a human model that possesses an optimal BBB phenotype and is readily scalable. To address this challenge, we developed a human in vitro BBB model comprising brain microvascular endothelial cells (BMECs), pericytes, astrocytes and neurons d...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Biological & Pharmaceutical Bulletin
سال: 2021
ISSN: ['1347-5215', '0918-6158']
DOI: https://doi.org/10.1248/bpb.b21-00218