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iCell Blood-Brain Barrier Isogenic Kit

Reliably replicate a functional blood-brain barrier (BBB) in vitro with an isogenic co-culture of human endothelial cells, astrocytes, and pericytes.

  • Superior integrity The iCell® blood-brain barrier assay kit demonstrates barrier properties comparable to those of the human BBB.
  • Versatile applications Long-term survival supports varied assays, including transporter function and compound permeability.
  • Fast recovery Consisting of terminally differentiated induced pluripotent stem cells (iPSCs), the BBB kit is ready from thaw.
  • Reproducible research FUJIFILM CDI’s iPSC expertise and rigorous quality control process ensure a consistent, scalable BBB model.
  • System compatibility Integrating the iCell BBB kit with organ-on-a-chip technologies replicates physiological complexity.
$4,200.00
Catalog # R1241
OUT OF STOCK
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Products

Kits

  • 3M, Cryopreserved iCell Brain Microvascular Endothelial Cells, 01279
  • 1.5M, Cryopreserved iCell Pericytes, 01279
  • 1M, Cryopreserved iCell Astrocytes 2.0, 01279
  • 200 µl, iCell Plating Supplement
  • 30 ml, iCell Astrocyte & Pericyte Medium
  • 100 ml, iCell BMEC Maintenance Medium

genetic status

quantity (Cells Per Vials)

catalog #

Catalog #: R1241

iCell Blood-Brain Barrier Isogenic Kit

From
$4,200.00
OUT OF STOCK

Product Overview

The blood-brain barrier (BBB) functions to maintain a tightly controlled microenvironment around the brain. For many years, the drug discovery market has needed a robust in vitro BBB model system to evaluate drug permeability and barrier function, as well as to study the diseases that affect it.

The inherent power of iPSC technology provides access to the specialized cell types required to assemble such a model system, but the field has been plagued with difficulties in reliably manufacturing a consistent supply of these cells at-scale. FUJIFILM Cellular Dynamics Inc., the market leader in iPSC technology and innovation, has developed a new human iPSC-derived BBB isogenic kit to model the BBB system in a cell culture insert format. The iCell Blood-Brain Barrier Isogenic Kit includes iCell Astrocytes 2.0, iCell Brain Microvascular Endothelial Cells (iCell BMEC) and iCell Pericytes, (all derived from the same proprietary donor background 01279) , and media that enables their long-term survival and superior functional performance assessed by transendothelial electrical resistance (TEER) assays.

The iCell Blood-Brain Barrier Isogenic Kit components have the potential to integrate with emerging organ-on-a-chip technologies and other 3D cell culture systems, thus offering an exciting new capability for the drug discovery community to advance the understanding of BBB function with respect to human health and disease. 

The iCell Blood-Brain Barrier Isogenic Kit:

  • Utilizes our newly developed cryopreserved iCell Brain Microvascular Endothelial Cells, iCell Pericytes, and iCell Astrocytes 2.0 generated from the same donor line (isogenic).
  • Includes our novel plating and co-culture medium.
  • Achieves superior transendothelial electrical resistance (TEER) values within 5 days.
  • Enables long-term studies.

Components:

Catalog #R1241
iCell Blood-Brain Barrier Isogenic Kit,
  • 3M, Cryopreserved iCell Brain Microvascular Endothelial Cells, 01279
  • 1.5M, Cryopreserved iCell Pericytes, 01279
  • 1M, Cryopreserved iCell Astrocytes 2.0, 01279
  • 200 µl, iCell Plating Supplement
  • 30 ml, iCell Astrocyte & Pericyte Medium
  • 100 ml, iCell BMEC Maintenance Medium

Donor Background

Sex:
Male
Race:
Caucasian
Cell Type:
Astrocytes; Brain Microvascular Endothelial Cells; Pericytes
Source Line:
01279
Modeled Disease Phenotype:
Apparently Healthy Normal
Donor Status:
Healthy
Tissue Source:
Blood
Age At Sampling:
50-59
Engineered Line:
No
Disease Status:
Apparently Healthy Normal (AHN)

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Need Help With This Product?

Our specialists are here to help you find the best product for your application.

Call Us

(877) 320-6688

 

OR

Our regular business hours are 9:00am to 5:00pm Central Time (USA)

Performance Data

Characterization of iPSC-derived Brain Microvascular Endothelial Cells

Brain microvascular endothelial cells (BMEC) are unlike other vascular endothelial cells lining peripheral blood vessels in that they display distinctive morphological, structural, and functional features. iCell® BMEC have cobblestone morphology as tightly packed cells with uniform size and clear cell boundaries. iCell BMEC marker expression (by ICC and flow cytometry) reveals characteristic endothelial markers (ZO-1, Claudin 5), transporters (GLUT1, CD98hc), and efflux/ influx proteins (BCRP, P-gp, MRP1, TfRc; these data not shown here). iCell BMEC exhibit much higher TEER functionality compared to iCell Endothelial Cells.

Characteristics of iPSC-derived iCell® Astrocytes 2.0

iCell Astrocytes 2.0 are Highly Pure Population of Human iPSC-derived Astrocytes. Top) iCell Astrocytes 2.0 were analyzed at thaw using flow cytometry, showing a highly pure population of astrocytes as identified by expression of CD44, GFAP, and CD49f. Bottom) Representative immunocytochemistry images of iCell Astrocytes 2.0 at 7 days post thaw showing expression of CD44, CD49f, S100b, and GFAP.

Characterization of iPSC-derived Pericytes

Pericytes are mural cells (like vascular smooth muscle) which ensheathe endothelium throughout the body. They are an important element of the BBB and function to maintain homeostasis of the barrier, regulate blood flow, and facilitate neuro-inflammatory responses. iCell® Pericytes display similar morphology to primary cells and express relevant markers (PDGFR-beta, NG2, CD13, and CD146) by ICC and flow cytometry. These cells also display functional phagocytic activity of S. aureus bioparticles in mono-culture (data not shown here).

TEER Assay with Human iPSC-derived iCell® BBB Isogenic Model

Transendothelial Electrical Resistance (TEER) is a widely accepted technique to measure barrier integrity and tight junction dynamics for a cell monolayer. (A) iCell BBB Isogenic Kit was developed and optimized using a sandwich assay on Transwell cell culture inserts. On Day (-2) of the TEER assay, the apical side of the microporous membrane is coated with fibronectin and collagen-IV, while the basolateral side is coated with gelatin. On Day (-1), iCell Astrocytes and iCell Pericytes are seeded together on the basolateral side in BBB Medium #1. On Day 0, iCell BMEC are thawed and plated on the apical side in BBB Medium #2. (B) Testing of the functional BBB tri-culture system is ready as soon as Day 2 with values typically beginning >500 Ω·cm2. Addition of multiple cell types increases the TEER values and reproducibility of the data (smaller error bars). (C) Timing of when to record TEER post-media change, as well as the formulation of assay media, can influence TEER values and should be considered before starting a complex experiment. Serum-free assay media can be substituted to completely minimize any interactions with test compounds while still maintaining physiologically-relevant barrier function. This experiment was performed on Day 5 after assembly of BBB tri-culture. Pre-Feed = TEER recording prior to any media change.

Receptor-Mediated Transcytosis assay using iCell® BBB Isogenic Kit

RMT is the primary transport route of large endogenous and exogenous molecules across the BBB into the central nervous system. As shown in the figure:

  • (A) transferrin crosses the cell culture insert in the absence of cells
  • (B) in the presence of the iCell BBB tri-culture, transferrin passes the BBB via RMT by binding to TfRs expressed by iCell BMECs
  • (C) the inhibitor ferrostatin II binds to TfRs, preventing transferrin transport across the BBB

OrganoPlate seeded with components of the iCell® BBB Isogenic Kit: TEER data

A & B. Brightfield images of single chips on an OrganoPlate of both mono-culture (A) and tri-culture (B) of the iCell BBB Isogenic Kit acquired with Spheroid setting on IncuCyte SX5. C. TEER measurements increase with the number of iCell BMECs seeded in the top channel of the device. 40,000 iCell BMECs resulted in the highest average TEER values. TEER measurements were made between days 2 and 5.

Immunostaining of iCell® BBB on OrganoPlate

Tri-culture of isogenic iCell BBB kit in one chip of a 40-chip 3-lane OrganoPlate. 40,000 iCell BMECs were plated in the Matrigel coated top channel. 40,000 iCell Pericytes and 20,000 iCell Astrocyte 2.0 were plated in the bottom gelatin coated channel. ZO-1 (red) in top channel indicates the formation of expected tight junctions between BMECs. GFAP stained Astrocytes (green) and PDGFR-β (pink) indicate the presence of relevant supporting cell types in the tri-culture.

Product Highlights

iCell Blood-Brain Barrier Isogenic Kit Highlights

  • Easy to implement - iCell Blood-Brain Barrier Isogenic Kits are shipped cryopreserved with optimized media. Simply thaw, plate and assay.
  • Human cells - The three cell types (iCell Brain Microvascular Endothelial Cells, iCell Pericytes, and iCell Astrocytes 2.0) in the iCell Blood-Brain Barrier Isogenic Kit are differentiated from human iPS cells from the same donor.
  • Superior function - iCell Blood-Brain Barrier Isogenic Kit in vitro model enables long-term survival and superior functional performance assessed by transendothelial electrical resistance (TEER) assay (>1500 ohm * cm2) in cell culture insert format.
  • Reproducible - Rigorous quality control ensures the same performance with every batch to yield reliable and reproducible results.
  • Compatible with Organ-On-Chip (OOC) platforms - The iCell Blood-Brain Barrier Isogenic Kits can be used with some optimization with many of the commercial OOC platforms.