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The Aggregate Induced Differentiation of Dopaminergic Neurons in NT2 Cells

Permanent Link: http://ncf.sobek.ufl.edu/NCFE003970/00001

Material Information

Title: The Aggregate Induced Differentiation of Dopaminergic Neurons in NT2 Cells Mediation by Transcription Factors Pax-2, Pitx3 and Otx2
Physical Description: Book
Language: English
Creator: McGrew, Christa
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2008
Publication Date: 2008

Subjects

Subjects / Keywords: Dopamine
Stem Cells
Parkinson's Disease
Transcription Factors
Differentiation
Central Nevous System
Neurodegenerative Disorders
Cell Replacement Therapy
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: The Ntera2 (NT2) cell line is a monoclonal, pluripotent line that shows strong similarities to the progenitor cells of the central nervous system (CNS). Aggregation in suspension culture induces differentiation of the NT2 line into neurons expressing a variety of phenotypes, astrocytes, and glial cells. Because of their rapid proliferation in vitro, NT2 cells have been targeted as a cell source for cell replacement treatments for traumatic brain injury, spinal cord injury, and a number of neurodegenerative disorders. Otx2, Pax-2, and Pitx3 are three transcription factors that direct the differentiation and maturation of dopaminergic neurons during in vivo midbrain development. This study uses siRNA gene knockdown, realtime RT-PCR, and immunocytochemistry (ICC) staining to explore the roles of Otx2, Pax-2, and Pitx3 during the in vitro, aggregate-induced differentiation of NT2 cells. siRNA gene knockdown of Pax-2 and Pitx3 during the aggregation stage of induction did not decrease the percent of cells expressing the dopaminergic cell marker tyrosine hydroxylase (TH). However, there was an observed morphological change in TH-positive cells in Pax-2 and Pitx3 siRNA knockdown as compared to control cell cultures. THpositive cells in control cultures grew in large clusters displaying extensive neurite outgrowth, while those within Pax-2 and Pitx3 siRNA cultures typically displayed a spherical morphology and did not grow in clusters. Real-time RT-PCR and ICC staining results showed an up-regulation of all three transcription factors in aggregate-induced, differentiated cell cultures as compared to the pre-differentiated, pluripotent cell cultures.
Statement of Responsibility: by Christa McGrew
Thesis: Thesis (B.A.) -- New College of Florida, 2008
Electronic Access: RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE
Bibliography: Includes bibliographical references.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Local: Faculty Sponsor: Walstrom, Katherine

Record Information

Source Institution: New College of Florida
Holding Location: New College of Florida
Rights Management: Applicable rights reserved.
Classification: local - S.T. 2008 M13
System ID: NCFE003970:00001

Permanent Link: http://ncf.sobek.ufl.edu/NCFE003970/00001

Material Information

Title: The Aggregate Induced Differentiation of Dopaminergic Neurons in NT2 Cells Mediation by Transcription Factors Pax-2, Pitx3 and Otx2
Physical Description: Book
Language: English
Creator: McGrew, Christa
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2008
Publication Date: 2008

Subjects

Subjects / Keywords: Dopamine
Stem Cells
Parkinson's Disease
Transcription Factors
Differentiation
Central Nevous System
Neurodegenerative Disorders
Cell Replacement Therapy
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: The Ntera2 (NT2) cell line is a monoclonal, pluripotent line that shows strong similarities to the progenitor cells of the central nervous system (CNS). Aggregation in suspension culture induces differentiation of the NT2 line into neurons expressing a variety of phenotypes, astrocytes, and glial cells. Because of their rapid proliferation in vitro, NT2 cells have been targeted as a cell source for cell replacement treatments for traumatic brain injury, spinal cord injury, and a number of neurodegenerative disorders. Otx2, Pax-2, and Pitx3 are three transcription factors that direct the differentiation and maturation of dopaminergic neurons during in vivo midbrain development. This study uses siRNA gene knockdown, realtime RT-PCR, and immunocytochemistry (ICC) staining to explore the roles of Otx2, Pax-2, and Pitx3 during the in vitro, aggregate-induced differentiation of NT2 cells. siRNA gene knockdown of Pax-2 and Pitx3 during the aggregation stage of induction did not decrease the percent of cells expressing the dopaminergic cell marker tyrosine hydroxylase (TH). However, there was an observed morphological change in TH-positive cells in Pax-2 and Pitx3 siRNA knockdown as compared to control cell cultures. THpositive cells in control cultures grew in large clusters displaying extensive neurite outgrowth, while those within Pax-2 and Pitx3 siRNA cultures typically displayed a spherical morphology and did not grow in clusters. Real-time RT-PCR and ICC staining results showed an up-regulation of all three transcription factors in aggregate-induced, differentiated cell cultures as compared to the pre-differentiated, pluripotent cell cultures.
Statement of Responsibility: by Christa McGrew
Thesis: Thesis (B.A.) -- New College of Florida, 2008
Electronic Access: RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE
Bibliography: Includes bibliographical references.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Local: Faculty Sponsor: Walstrom, Katherine

Record Information

Source Institution: New College of Florida
Holding Location: New College of Florida
Rights Management: Applicable rights reserved.
Classification: local - S.T. 2008 M13
System ID: NCFE003970:00001

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