Effects of Collagen Coating, Fetal Bovine Serum Concentration, Differentiation Agents, and Neurotoxin Application on In Vitro Modeling of Parkinson's Disease Using SH-SY5Y Cell Culture

Fatma Gonca Koçancı

Makale detayları

Effects of Collagen Coating, Fetal Bovine Serum Concentration, Differentiation Agents, and Neurotoxin Application on In Vitro Modeling of Parkinson's Disease Using SH-SY5Y Cell Culture

Yazarlar:

Fatma Gonca Koçancı

Yayıncı:

Alanya Alaaddin Keykubat Üniversitesi

Ana Başlıklar:

Geriatri ve Gerontoloji

Dergi:

Acta Medica Alanya Vol.8 Issue 2, pp.105-109

Belge Tipi:

Makale

Yayın Tarihi:

8/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001326051

pISSN:

2587-0319

Anahtar kelimeler:

Özet:

 Aim: This study aims to optimize SH-SY5Y culture conditions to develop precise in vitro disease models for Parkinsons disease PD research. It seeks to investigate the effects of various factors such as collagen coating, fetal bovine serum concentration, differentiation agents, and neurotoxin treatments on cellular behavior and disease modeling. Materials and Methods: The human neuroblastoma SH-SY5Y cell line was cultured in DMEMF12 supplemented with heat-inactivated fetal bovine serum FBS, penicillin-streptomycin, and L-glutamine. Collagen coating was applied to assess its impact on cell differentiation, while the ideal cell density and serum ratio for generating neurite-like cells were determined through experimentation. The MTT assay was employed to evaluate the cytotoxic effects of paraquat, while dopamine levels were quantified using ELISA. Gene expression was analyzed via real-time qPCR. Immunofluorescence staining and neurite length measurements were conducted to validate the PD model and assess cellular morphology. Results: Cells cultured at a density of 5x103 cellscm2 with collagen and 2 FBS exhibited characteristics of dopaminergic neurons upon exposure to retinoic acid. Conversely, paraquat treatment induced neurotoxicity, resulting in decreased dopamine levels and impaired neurite outgrowth. Conclusion: This study investigated the optimization of SH-SY5Y cell culture conditions for PD modeling. Key findings include optimal cell density, FBS concentration, and beneficial effects of collagen coating. Additionally, an effective paraquat neurotoxicity protocol has been established, providing a solid framework for future research on neuronal differentiation and degeneration.

 Aim: This study aims to optimize SH-SY5Y culture conditions to develop precise in vitro disease models for Parkinsons disease PD research. It seeks to investigate the effects of various factors such as collagen coating, fetal bovine serum concentration, differentiation agents, and neurotoxin treatments on cellular behavior and disease modeling. Materials and Methods: The human neuroblastoma SH-SY5Y cell line was cultured in DMEMF12 supplemented with heat-inactivated fetal bovine serum FBS, penicillin-streptomycin, and L-glutamine. Collagen coating was applied to assess its impact on cell differentiation, while the ideal cell density and serum ratio for generating neurite-like cells were determined through experimentation. The MTT assay was employed to evaluate the cytotoxic effects of paraquat, while dopamine levels were quantified using ELISA. Gene expression was analyzed via real-time qPCR. Immunofluorescence staining and neurite length measurements were conducted to validate the PD model and assess cellular morphology. Results: Cells cultured at a density of 5x103 cellscm2 with collagen and 2 FBS exhibited characteristics of dopaminergic neurons upon exposure to retinoic acid. Conversely, paraquat treatment induced neurotoxicity, resulting in decreased dopamine levels and impaired neurite outgrowth. Conclusion: This study investigated the optimization of SH-SY5Y cell culture conditions for PD modeling. Key findings include optimal cell density, FBS concentration, and beneficial effects of collagen coating. Additionally, an effective paraquat neurotoxicity protocol has been established, providing a solid framework for future research on neuronal differentiation and degeneration.

 Aim: This study aims to optimize SH-SY5Y culture conditions to develop precise in vitro disease models for Parkinsons disease PD research. It seeks to investigate the effects of various factors such as collagen coating, fetal bovine serum concentration, differentiation agents, and neurotoxin treatments on cellular behavior and disease modeling. Materials and Methods: The human neuroblastoma SH-SY5Y cell line was cultured in DMEMF12 supplemented with heat-inactivated fetal bovine serum FBS, penicillin-streptomycin, and L-glutamine. Collagen coating was applied to assess its impact on cell differentiation, while the ideal cell density and serum ratio for generating neurite-like cells were determined through experimentation. The MTT assay was employed to evaluate the cytotoxic effects of paraquat, while dopamine levels were quantified using ELISA. Gene expression was analyzed via real-time qPCR. Immunofluorescence staining and neurite length measurements were conducted to validate the PD model and assess cellular morphology. Results: Cells cultured at a density of 5x103 cellscm2 with collagen and 2 FBS exhibited characteristics of dopaminergic neurons upon exposure to retinoic acid. Conversely, paraquat treatment induced neurotoxicity, resulting in decreased dopamine levels and impaired neurite outgrowth. Conclusion: This study investigated the optimization of SH-SY5Y cell culture conditions for PD modeling. Key findings include optimal cell density, FBS concentration, and beneficial effects of collagen coating. Additionally, an effective paraquat neurotoxicity protocol has been established, providing a solid framework for future research on neuronal differentiation and degeneration.

 Amaç: Bu çalışma, Parkinson hastalığı PH araştırmaları için doğru in vitro hastalık modelleri geliştirmek amacıyla SH-SY5Y kültür koşullarını optimize etmeyi amaçlamaktadır. Kollajen kaplama, fetal sığır serum FSS konsantrasyonu, diferansiyasyon ajanları ve nörotoksin tedavileri gibi çeşitli faktörlerin hücresel davranış ve hastalık modellemesi üzerindeki etkilerini araştırmayı hedeflemektedir. Yöntem: İnsan nöroblastoma SH-SY5Y hücre hattı, ısı ile inaktive edilmiş fetal sığır serumu, penisilin-streptomisin ve L-glutamin ile desteklenmiş DMEMF12 içinde kültürlenmiştir. Hücre diferansiyasyonu üzerindeki etkisini değerlendirmek için kollajen kaplama uygulanmış, ideal hücre yoğunluğu ve serum oranı ise deneysel olarak belirlenmiştir. Parakuatın sitotoksik etkilerini değerlendirmek için MTT testi kullanılmış, dopamin seviyeleri ELISA ile ölçülmüştür. Gen ekspresyonu gerçek zamanlı qPCR ile analiz edilmiştir. Parkinson modelini doğrulamak ve hücresel morfolojiyi değerlendirmek için immünofloresan boyama ve nörit uzunluğu ölçümleri yapılmıştır. Bulgular: 5x103 hücrecm2 yoğunluğunda kültürlenmiş hücreler, kollajen ve 2 FSS ile retinoik asit maruziyetinde dopaminerjik nöron özellikleri sergilemiştir. Bununla birlikte, parakuat tedavisi nörotoksisiteye neden olmuş, dopamin seviyelerinde azalma ve nörit büyümesinde bozulma gözlenmiştir. Sonuç: Bu çalışma, PH modellemesi için SH-SY5Y hücre kültürü koşullarınının optimizasyonu araştırmıştır. Temel bulgular arasında optimal hücre yoğunluğu, FSS konsantrasyonu ve kolajen kaplamanın faydalı etkileri yer almaktadır. Ek olarak, nöronal farklılaşma ve dejenerasyon konusunda gelecekteki araştırmalar için sağlam bir çerçeve sağlayan etkili bir parakuat nörotoksisite protokolü oluşturulmuştur.

Makale detayları

Effects of Collagen Coating, Fetal Bovine Serum Concentration, Differentiation Agents, and Neurotoxin Application on In Vitro Modeling of Parkinson's Disease Using SH-SY5Y Cell Culture

Yazarlar:

Fatma Gonca Koçancı

Yayıncı:

Alanya Alaaddin Keykubat Üniversitesi

Ana Başlıklar:

Geriatri ve Gerontoloji

Dergi:

Acta Medica Alanya Vol.8 Issue 2, pp.105-109

Belge Tipi:

Makale

Yayın Tarihi:

8/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001326051

pISSN:

2587-0319

Parkinson

SH-SY5Y cells

neuronal differentiation

neurotoxicity

in vitro modelingParkinson hastalığı

SH-SY5Y hücreleri

nöronal diferansiyasyon

nörotoksisite

in vitro modelleme

Sayı

Makale detayları

Yazarlar: Fatma Gonca Koçancı

Yayıncı: Alanya Alaaddin Keykubat Üniversitesi

Ana Başlıklar: Geriatri ve Gerontoloji

Dergi: Acta Medica Alanya Vol.8 Issue 2, pp.105-109

Belge Tipi: Makale

Yayın Tarihi: 8/2024

Belge Dili: İngilizce

Document ID (Org No.): 2001326051

pISSN: 2587-0319

Sayı

Yazarlar:

Fatma Gonca Koçancı

Yayıncı:

Alanya Alaaddin Keykubat Üniversitesi

Ana Başlıklar:

Geriatri ve Gerontoloji

Dergi:

Acta Medica Alanya Vol.8 Issue 2, pp.105-109

Belge Tipi:

Makale

Yayın Tarihi:

8/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001326051

pISSN:

2587-0319