Cell Delivery System for Traumatic Brain Injury

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Cell Delivery System for Traumatic Brain Injury

• W. Wiesmann M.D.1, M. Sarvestani2, J.
  Manassero3, J. Kim3, M. St Georges3, N.
  Esclamado3, I. Wulur1, E. Orwin2,3
• 1BioSTAR, Inc.
• 2BioSTAR West, Claremont, Ca
• 3Harvey Mudd College, Claremont, Ca

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Traumatic Brain Injury (TBI)

• Background info on TBI
• Our approach Deliver high density cells in a
  scaffold

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What is the Microenvironment Necessary to Restore
Function after TBI?

• Differentiation state of cells (human bone
  marrow-derived stem cells, MSCs)
• Appropriate scaffold material and configuration
• Stimulatory growth factors

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Experimental Plan

• Cells
• MSCs alone
• MSCs differentiated neurons via retinoic
  acid (RA)
• MSCs differentiated neurons via co-culture
  with astrocytes
• Scaffold
• Collagen
• Chitosan
• Collagen/chitosan hybrid
• Scaffold structure hydrogel vs. sponge vs.
  electrospun tube

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Results Cell Morphology Retinoic Acid

Day 14
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Results Cell Morphology RA Astrocytes
Day 14

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Results Cell Differentiation in Monolayer at Day
8
Blue MAP2 Red ß tubulin Green
Sytox mature neuron marker Intermdiate to late
neuron marker Nuclear stain
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Results Cell Differentiation of RA Treated MSCs
in Monolayer at Day 8
Blue MAP2 Red Synaptophysin Green
Sytox mature neuron marker Synaptic vesicle
associated protein Nuclear stain
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Preliminary Results Cell Differentiation in 3D
ScaffoldCo-culture with Astrocytes 14 days
A
B
A
Blue MAP2 Red ß tubulin Green
Sytox mature neuron marker Intermdiate to late
neuron marker Nuclear stain
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Preliminary Results Cell Differentiation in 3D
ScaffoldRetinoic Acid 12 days
Double click the images to play movie
Blue MAP2 Red Synaptophysin Green
Sytox mature neuron marker Synaptic vesicle
associated protein Nuclear stain
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Neural Project Status

• Positive MAP2 Synaptophysin expression on day 8
  samples grown in monolayer with 30 µM Retinoic
  Acid treatment
• Positive MAP2 expression on day 12 samples grown
  on collagen sponges with 30 µM Retinoic Acid
  treatment
• Synaptophysin is yet to be expressed on 12 day
  collagen samples