Astrocyte Growth and Glial Cell Line-Derived Neurotrophic Factor Secretion in Three-Dimensional Polyethylene Terephthalate Fibrous Matrices

Abstract

The ability of human astrocytes grown in nonwoven fibrous matrices to produce glial cell line-derived neurotrophic factor (GDNF) was studied. GDNF has the ability to selectively nourish and regenerate dopaminergic neurons and thus can provide a new treatment of Parkinson's disease. Compressed polyethylene terephthalate (PET) fabrics (porosity, 88.8%; mean pore diameter, 64 microm), treated with boiling NaOH, was effective in supporting high-density growth of astrocytes with stable GDNF production over the entire period of 18 days studied. Treatment of PET with NaOH renders the fiber surface more hydrophilic, thereby facilitating attachment and spreading of cells, whereas matrix compression allows cells to grow along and also between the fibers of these matrices to a higher density. The average production of GDNF by cells grown in these matrices (approximately 2 cm in diameter) was 21.7 pg/mL x day, with an average high concentration of 64.6 pg/mL, which is well above the effective concentration of 40 pg/mL. This work shows promise in culturing astrocytes in PET matrices as the first step in developing a potential implantable tissue-engineering device for treating patients with Parkinson's disease.

Influence of Probe Flexibility and Gelatin Embedding on Neuronal Density and Glial Responses to Brain Implants

“…Importantly, no obvious loss of neurons in the inner ROIs for the gelatin embedded probes were found, whereas there was a reduced survival of neurons in the inner ROIs of the non-embedded probes, as previously reported [ 19 ]. These results support previous findings suggesting that gelatin embedding is beneficial for neuronal survival [ 26 , 27 ]. It should also be noticed that neurons in the inner ROI appeared less deformed when implants were allowed to flex in the principal direction of movement within the brain parenchyma ( Fig.…”

Influence of Probe Flexibility and Gelatin Embedding on Neuronal Density and Glial Responses to Brain Implants

“…Importantly, no obvious loss of neurons in the inner ROIs for the gelatin embedded probes were found, whereas there was a reduced survival of neurons in the inner ROIs of the non-embedded probes, as previously reported [ 19 ]. These results support previous findings suggesting that gelatin embedding is beneficial for neuronal survival [ 26 , 27 ]. It should also be noticed that neurons in the inner ROI appeared less deformed when implants were allowed to flex in the principal direction of movement within the brain parenchyma ( Fig.…”

Benzalkonium Chloride Sterilization of Nonwoven Fibrous Scaffolds for Astrocyte Culture

“…Astrocytes play a significant role in the central nervous system. It has been previously suggested that astrocytes cultured in PET scaffolds could provide a transplantable cell therapy platform for treating the Parkinson's disease [29]. In this work, long-term astrocyte cultures in PET scaffolds were demonstrated to confirm the suitability and effectiveness of the BAC sterilization method.…”

Two‐color fluorescent proteins reporting survivin regulation in breast cancer cells for high throughput drug screening

“…Using the fluorescent signal directly as the reporter can also minimize errors due to large variations in the spheroid size in 3D cultures using conventional viability assays and microscopic image analysis (Zanoni et al, 2016), which is cumbersome to use in high throughout screening (Booij et al, 2019). However, 3D scaffolds, such as the nonwoven fibrous PET matrix, used in cell‐based assays must be optimized in its macro‐ and micro‐structures, including pore size and surface properties, which have profound effects on cell proliferation, differentiation, and function (Basu & Yang, 2005; Ma et al, 1999; 2000; Y. Yang et al, 2005), and thus can affect cell responses to drugs (Zang et al, 2016).…”

Influence of Probe Flexibility and Gelatin Embedding on Neuronal Density and Glial Responses to Brain Implants

“…Importantly, no obvious loss of neurons in the inner ROIs for the gelatin embedded probes were found, whereas there was a reduced survival of neurons in the inner ROIs of the non-embedded probes, as previously reported [ 19 ]. These results support previous findings suggesting that gelatin embedding is beneficial for neuronal survival [ 26 , 27 ]. It should also be noticed that neurons in the inner ROI appeared less deformed when implants were allowed to flex in the principal direction of movement within the brain parenchyma ( Fig.…”

Benzalkonium Chloride Sterilization of Nonwoven Fibrous Scaffolds for Astrocyte Culture

“…Astrocytes play a significant role in the central nervous system. It has been previously suggested that astrocytes cultured in PET scaffolds could provide a transplantable cell therapy platform for treating the Parkinson's disease [29]. In this work, long-term astrocyte cultures in PET scaffolds were demonstrated to confirm the suitability and effectiveness of the BAC sterilization method.…”

Two‐color fluorescent proteins reporting survivin regulation in breast cancer cells for high throughput drug screening

“…Using the fluorescent signal directly as the reporter can also minimize errors due to large variations in the spheroid size in 3D cultures using conventional viability assays and microscopic image analysis (Zanoni et al, 2016), which is cumbersome to use in high throughout screening (Booij et al, 2019). However, 3D scaffolds, such as the nonwoven fibrous PET matrix, used in cell‐based assays must be optimized in its macro‐ and micro‐structures, including pore size and surface properties, which have profound effects on cell proliferation, differentiation, and function (Basu & Yang, 2005; Ma et al, 1999; 2000; Y. Yang et al, 2005), and thus can affect cell responses to drugs (Zang et al, 2016).…”

Influence of Probe Flexibility and Gelatin Embedding on Neuronal Density and Glial Responses to Brain Implants

“…Importantly, no obvious loss of neurons in the inner ROIs for the gelatin embedded probes were found, whereas there was a reduced survival of neurons in the inner ROIs of the non-embedded probes, as previously reported [ 19 ]. These results support previous findings suggesting that gelatin embedding is beneficial for neuronal survival [ 26 , 27 ]. It should also be noticed that neurons in the inner ROI appeared less deformed when implants were allowed to flex in the principal direction of movement within the brain parenchyma ( Fig.…”

Benzalkonium Chloride Sterilization of Nonwoven Fibrous Scaffolds for Astrocyte Culture

“…Astrocytes play a significant role in the central nervous system. It has been previously suggested that astrocytes cultured in PET scaffolds could provide a transplantable cell therapy platform for treating the Parkinson's disease [29]. In this work, long-term astrocyte cultures in PET scaffolds were demonstrated to confirm the suitability and effectiveness of the BAC sterilization method.…”

Two‐color fluorescent proteins reporting survivin regulation in breast cancer cells for high throughput drug screening

“…Using the fluorescent signal directly as the reporter can also minimize errors due to large variations in the spheroid size in 3D cultures using conventional viability assays and microscopic image analysis (Zanoni et al, 2016), which is cumbersome to use in high throughout screening (Booij et al, 2019). However, 3D scaffolds, such as the nonwoven fibrous PET matrix, used in cell‐based assays must be optimized in its macro‐ and micro‐structures, including pore size and surface properties, which have profound effects on cell proliferation, differentiation, and function (Basu & Yang, 2005; Ma et al, 1999; 2000; Y. Yang et al, 2005), and thus can affect cell responses to drugs (Zang et al, 2016).…”