Establishment of three-dimensional culture of neural stem/progenitor cells in collagen Type-1 Gel

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Abstract

Purpose: Recent studies demonstrated that transplanting neural stem/precursor cells (NSPCs) into an injured spinal cord of adult rat promotes functional recovery. The functional recovery from spinal cord injury might be enhanced by transplanting NSPCs with a scaffold that fills the cavity, entraps the NSPCs in the cavity, and acts as an attachment for neurite extension. We recently focused on collagen type-1 as a scaffold for NSPC transplantation into the injured spinal cord. In the present study, we determined the optimal conditions for culturing NSPCs in 3D collagen type-1 gel with respect to cell survival and cell migration. We, then, evaluated the ability of NSPCs to differentiate under the optimal condition. Methods: NSPCs were derived from the striatum of rat embryos. To determine the optimal cell density and collagen concentration for 3D collagen gel culture for NSPC, we performed viability assay and migration assay. Then, we examined the proportion of phenotypes differentiated from NSPC in that optimal condition. Results: In viability assay, the viability rate increased as the NSPC density increased, and peaked at 1 × 107 to 5 × 107 cell/ml. For collagen concentration, the viability rate increased as the collagen concentration decreased. In migration assay, cell migration was most extensive at collagen concentrations between 0.3 and 0.75 mg/ml. Migration distances gradually declined as collagen concentrations increased. In the optimal condition, NSPCs differentiated into neurons (40.1%), astrocytes (53.1%), and oligodendrocytes (5.3%) in 3D collagen gel culture. Conclusion: The optimal conditions for NSPC culture in 3D collagen gel was a cell density between 1 × 107 and 5 × 107 cells/ml and a collagen concentration between 0.5 and 0.75 mg/ml. Under the condition, NSPCs could differentiate into neurons, astrocytes, and oligodendrocytes.

Original languageEnglish
Pages (from-to)109-117
Number of pages9
JournalRestorative Neurology and Neuroscience
Volume25
Issue number2
Publication statusPublished - 2007

Fingerprint

Neural Stem Cells
Collagen Type I
Stem Cells
Gels
Collagen
Cell Count
Oligodendroglia
Astrocytes
Cell Movement
Cell Survival
Spinal Cord
Neurons
Stem Cell Transplantation
Neurites
Spinal Cord Injuries
Embryonic Structures
Cell Culture Techniques

Keywords

  • Collagen type-1
  • Neural stem precursor cell
  • Scaffold
  • Three-dimensional culture

ASJC Scopus subject areas

  • Neuroscience(all)
  • Neuropsychology and Physiological Psychology

Cite this

@article{6e1ff69dd7f642f79eb35d8aa25fd63d,
title = "Establishment of three-dimensional culture of neural stem/progenitor cells in collagen Type-1 Gel",
abstract = "Purpose: Recent studies demonstrated that transplanting neural stem/precursor cells (NSPCs) into an injured spinal cord of adult rat promotes functional recovery. The functional recovery from spinal cord injury might be enhanced by transplanting NSPCs with a scaffold that fills the cavity, entraps the NSPCs in the cavity, and acts as an attachment for neurite extension. We recently focused on collagen type-1 as a scaffold for NSPC transplantation into the injured spinal cord. In the present study, we determined the optimal conditions for culturing NSPCs in 3D collagen type-1 gel with respect to cell survival and cell migration. We, then, evaluated the ability of NSPCs to differentiate under the optimal condition. Methods: NSPCs were derived from the striatum of rat embryos. To determine the optimal cell density and collagen concentration for 3D collagen gel culture for NSPC, we performed viability assay and migration assay. Then, we examined the proportion of phenotypes differentiated from NSPC in that optimal condition. Results: In viability assay, the viability rate increased as the NSPC density increased, and peaked at 1 × 107 to 5 × 107 cell/ml. For collagen concentration, the viability rate increased as the collagen concentration decreased. In migration assay, cell migration was most extensive at collagen concentrations between 0.3 and 0.75 mg/ml. Migration distances gradually declined as collagen concentrations increased. In the optimal condition, NSPCs differentiated into neurons (40.1{\%}), astrocytes (53.1{\%}), and oligodendrocytes (5.3{\%}) in 3D collagen gel culture. Conclusion: The optimal conditions for NSPC culture in 3D collagen gel was a cell density between 1 × 107 and 5 × 107 cells/ml and a collagen concentration between 0.5 and 0.75 mg/ml. Under the condition, NSPCs could differentiate into neurons, astrocytes, and oligodendrocytes.",
keywords = "Collagen type-1, Neural stem precursor cell, Scaffold, Three-dimensional culture",
author = "Koota Watanabe and Masaya Nakamura and Hideyuki Okano and Yoshiaki Toyama",
year = "2007",
language = "English",
volume = "25",
pages = "109--117",
journal = "Restorative Neurology and Neuroscience",
issn = "0922-6028",
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T1 - Establishment of three-dimensional culture of neural stem/progenitor cells in collagen Type-1 Gel

AU - Watanabe, Koota

AU - Nakamura, Masaya

AU - Okano, Hideyuki

AU - Toyama, Yoshiaki

PY - 2007

Y1 - 2007

N2 - Purpose: Recent studies demonstrated that transplanting neural stem/precursor cells (NSPCs) into an injured spinal cord of adult rat promotes functional recovery. The functional recovery from spinal cord injury might be enhanced by transplanting NSPCs with a scaffold that fills the cavity, entraps the NSPCs in the cavity, and acts as an attachment for neurite extension. We recently focused on collagen type-1 as a scaffold for NSPC transplantation into the injured spinal cord. In the present study, we determined the optimal conditions for culturing NSPCs in 3D collagen type-1 gel with respect to cell survival and cell migration. We, then, evaluated the ability of NSPCs to differentiate under the optimal condition. Methods: NSPCs were derived from the striatum of rat embryos. To determine the optimal cell density and collagen concentration for 3D collagen gel culture for NSPC, we performed viability assay and migration assay. Then, we examined the proportion of phenotypes differentiated from NSPC in that optimal condition. Results: In viability assay, the viability rate increased as the NSPC density increased, and peaked at 1 × 107 to 5 × 107 cell/ml. For collagen concentration, the viability rate increased as the collagen concentration decreased. In migration assay, cell migration was most extensive at collagen concentrations between 0.3 and 0.75 mg/ml. Migration distances gradually declined as collagen concentrations increased. In the optimal condition, NSPCs differentiated into neurons (40.1%), astrocytes (53.1%), and oligodendrocytes (5.3%) in 3D collagen gel culture. Conclusion: The optimal conditions for NSPC culture in 3D collagen gel was a cell density between 1 × 107 and 5 × 107 cells/ml and a collagen concentration between 0.5 and 0.75 mg/ml. Under the condition, NSPCs could differentiate into neurons, astrocytes, and oligodendrocytes.

AB - Purpose: Recent studies demonstrated that transplanting neural stem/precursor cells (NSPCs) into an injured spinal cord of adult rat promotes functional recovery. The functional recovery from spinal cord injury might be enhanced by transplanting NSPCs with a scaffold that fills the cavity, entraps the NSPCs in the cavity, and acts as an attachment for neurite extension. We recently focused on collagen type-1 as a scaffold for NSPC transplantation into the injured spinal cord. In the present study, we determined the optimal conditions for culturing NSPCs in 3D collagen type-1 gel with respect to cell survival and cell migration. We, then, evaluated the ability of NSPCs to differentiate under the optimal condition. Methods: NSPCs were derived from the striatum of rat embryos. To determine the optimal cell density and collagen concentration for 3D collagen gel culture for NSPC, we performed viability assay and migration assay. Then, we examined the proportion of phenotypes differentiated from NSPC in that optimal condition. Results: In viability assay, the viability rate increased as the NSPC density increased, and peaked at 1 × 107 to 5 × 107 cell/ml. For collagen concentration, the viability rate increased as the collagen concentration decreased. In migration assay, cell migration was most extensive at collagen concentrations between 0.3 and 0.75 mg/ml. Migration distances gradually declined as collagen concentrations increased. In the optimal condition, NSPCs differentiated into neurons (40.1%), astrocytes (53.1%), and oligodendrocytes (5.3%) in 3D collagen gel culture. Conclusion: The optimal conditions for NSPC culture in 3D collagen gel was a cell density between 1 × 107 and 5 × 107 cells/ml and a collagen concentration between 0.5 and 0.75 mg/ml. Under the condition, NSPCs could differentiate into neurons, astrocytes, and oligodendrocytes.

KW - Collagen type-1

KW - Neural stem precursor cell

KW - Scaffold

KW - Three-dimensional culture

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VL - 25

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JO - Restorative Neurology and Neuroscience

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