Creating An Effective Model with In Vitro Adipocyte Cell Culture for Obesity Research: Preliminary Study

Ezgi Nur Çil; Yasemin Soysal

Makale detayları

Creating An Effective Model with In Vitro Adipocyte Cell Culture for Obesity Research: Preliminary Study

Yazarlar:

Ezgi Nur Çil | Yasemin Soysal

Yayıncı:

Zonguldak Bulent Ecevit University

Ana Başlıklar:

Genetik ve Kişiselleştirilmiş Beslenme Bilimi | Gıda ve Beslenme Dengesi | Hücre Fizyolojisi

Dergi:

Türkiye Diyabet ve Obezite Dergisi Vol.8 Issue 1, pp.59-64

Belge Tipi:

Makale

Yayın Tarihi:

4/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001316563

pISSN:

2587-0335

Anahtar kelimeler:

Özet:

 Aim: The differentiation efficiency of 3T3-L1 preadipocytes is a crucial factor affecting studies on cellular mechanisms associated with obesity, diabetes and related disorders. Differentiation of 3T3-L1 cells is usually induced by an Adipogenic cocktail containing insulin, dexamethasone DEX, and 3-isobutyl-1-methylxanthine IBMX. There is a process that includes important points to consider when growing 3T3-L1 cells. It is a sensitive cell group and may löse its ability to differentiate in later passages. To solve this problem, for the purpose of our study, the differentiation efficiency of three different commonly used Adipogenic cocktails was compared. Additionally, the differentiation abilities of 3T3-L1 cells were tested by changing some parameters. Material and Methods: In our study, 3T3-L1 cell culture, differentiation of fibroblast cells into mature adipocyte cells, demonstration of lipids under the microscope with Oil Red O staining, and spectrophotometric measurement of the dye for quantitative analysis were used. Results: Experimental results Showed that the Adipogenic cocktail containing 0.5 mM IBMX, 0.25 μM DEX and 1 μgmL insülin was the most effective cocktail for 3T3-L1 cell differentiation p0.0001 and the effectiveness of differentiation was closely related to the concentrations of adipogenic agents. Conclusion: In our study, important modifications that can be used for the in vitro preclinical obesity model have been reported as an alternative to the protocol given by ATCC for 3T3-L1 cells.

 Aim: The differentiation efficiency of 3T3-L1 preadipocytes is a crucial factor affecting studies on cellular mechanisms associated with obesity, diabetes and related disorders. Differentiation of 3T3-L1 cells is usually induced by an Adipogenic cocktail containing insulin, dexamethasone DEX, and 3-isobutyl-1-methylxanthine IBMX. There is a process that includes important points to consider when growing 3T3-L1 cells. It is a sensitive cell group and may löse its ability to differentiate in later passages. To solve this problem, for the purpose of our study, the differentiation efficiency of three different commonly used Adipogenic cocktails was compared. Additionally, the differentiation abilities of 3T3-L1 cells were tested by changing some parameters. Material and Methods: In our study, 3T3-L1 cell culture, differentiation of fibroblast cells into mature adipocyte cells, demonstration of lipids under the microscope with Oil Red O staining, and spectrophotometric measurement of the dye for quantitative analysis were used. Results: Experimental results Showed that the Adipogenic cocktail containing 0.5 mM IBMX, 0.25 μM DEX and 1 μgmL insülin was the most effective cocktail for 3T3-L1 cell differentiation p0.0001 and the effectiveness of differentiation was closely related to the concentrations of adipogenic agents. Conclusion: In our study, important modifications that can be used for the in vitro preclinical obesity model have been reported as an alternative to the protocol given by ATCC for 3T3-L1 cells.

 Aim: The differentiation efficiency of 3T3-L1 preadipocytes is a crucial factor affecting studies on cellular mechanisms associated with obesity, diabetes and related disorders. Differentiation of 3T3-L1 cells is usually induced by an Adipogenic cocktail containing insulin, dexamethasone DEX, and 3-isobutyl-1-methylxanthine IBMX. There is a process that includes important points to consider when growing 3T3-L1 cells. It is a sensitive cell group and may löse its ability to differentiate in later passages. To solve this problem, for the purpose of our study, the differentiation efficiency of three different commonly used Adipogenic cocktails was compared. Additionally, the differentiation abilities of 3T3-L1 cells were tested by changing some parameters. Material and Methods: In our study, 3T3-L1 cell culture, differentiation of fibroblast cells into mature adipocyte cells, demonstration of lipids under the microscope with Oil Red O staining, and spectrophotometric measurement of the dye for quantitative analysis were used. Results: Experimental results Showed that the Adipogenic cocktail containing 0.5 mM IBMX, 0.25 μM DEX and 1 μgmL insülin was the most effective cocktail for 3T3-L1 cell differentiation p0.0001 and the effectiveness of differentiation was closely related to the concentrations of adipogenic agents. Conclusion: In our study, important modifications that can be used for the in vitro preclinical obesity model have been reported as an alternative to the protocol given by ATCC for 3T3-L1 cells.

 Amaç: 3T3-L1 preadipositlerin farklılaşma etkinliği obezite, diyabet ve ilgili bozukluklarla ilişkili hücresel mekanizmalar üzerine yapılan çalışmaları etkileyen önemli bir faktördür. 3T3-L1 hücrelerinin farklılaşması genellikle insülin, deksametazon DEX ve 3-izobütil- 1-metilksantin IBMX içeren bir adipojenik kokteyl ile indüklenmektedir. 3T3-L1 hücreleri çoğaltılırken dikkat edilmesi gereken önemli noktalar içeren bir sürece sahip hassas bir hücre grubu olup ilerleyen pasajlarda farklılaşma yeteneğini kaybedebilmektedir. Bu sorunu çözmek için, çalışmamızın amacı doğrultusunda, yaygın olarak kullanılan üç farklı adipojenik kokteylin farklılaşma verimliliği karşılaştırılmıştır. Bununla birlikte, birtakım parametreler değiştirilerek 3T3-L1 hücrelerinin farklılaşma yetenekleri test edilmiştir. Gereç ve Yöntemler: Çalışmamızda 3T3-L1 hücre kültürü, fibroblast hücrelerin olgun adiposit hücrelere farklılaştırılması, Oil Red O boyama ile lipitlerin mikroskop altında gösterilmesi ve kantitatif analiz için boyanın spektrofotometrik ölçümü yöntemleri kullanılmıştır. Bulgular: Deney sonuçları, 0,5 mM IBMX, 0,25 μM DEX ve 1 μgmL insülin içeren adipogenik kokteylin 3T3-L1 hücre farklılaşması için en etkili kokteyl olduğunu p0,0001 ve farklılaşmanın etkinliğinin adipojenik ajanların konsantrasyonları ile yakından ilişkili olduğunu göstermiştir. Sonuç: Çalışmamızda ATCCnin 3T3-L1 hücreleri için vermiş olduğu protokolün alternatifi olarak in vitro preklinik obezite modeli için kullanılabilecek önemli modifikasyonlar bildirilmiştir.

Makale detayları

Creating An Effective Model with In Vitro Adipocyte Cell Culture for Obesity Research: Preliminary Study

Yazarlar:

Ezgi Nur Çil | Yasemin Soysal

Yayıncı:

Zonguldak Bulent Ecevit University

Ana Başlıklar:

Genetik ve Kişiselleştirilmiş Beslenme Bilimi | Gıda ve Beslenme Dengesi | Hücre Fizyolojisi

Dergi:

Türkiye Diyabet ve Obezite Dergisi Vol.8 Issue 1, pp.59-64

Belge Tipi:

Makale

Yayın Tarihi:

4/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001316563

pISSN:

2587-0335

Obesity

cell culture

3T3-L1

adipogenesis

adipocyteObezite

hücre kültürü

3T3-L1

adipogenez

adiposit

Sayı

Makale detayları

Yazarlar: Ezgi Nur Çil | Yasemin Soysal

Yayıncı: Zonguldak Bulent Ecevit University

Ana Başlıklar: Genetik ve Kişiselleştirilmiş Beslenme Bilimi | Gıda ve Beslenme Dengesi | Hücre Fizyolojisi

Dergi: Türkiye Diyabet ve Obezite Dergisi Vol.8 Issue 1, pp.59-64

Belge Tipi: Makale

Yayın Tarihi: 4/2024

Belge Dili: Türkçe

Document ID (Org No.): 2001316563

pISSN: 2587-0335

Sayı

Yazarlar:

Ezgi Nur Çil | Yasemin Soysal

Yayıncı:

Zonguldak Bulent Ecevit University

Ana Başlıklar:

Genetik ve Kişiselleştirilmiş Beslenme Bilimi | Gıda ve Beslenme Dengesi | Hücre Fizyolojisi

Dergi:

Türkiye Diyabet ve Obezite Dergisi Vol.8 Issue 1, pp.59-64

Belge Tipi:

Makale

Yayın Tarihi:

4/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001316563

pISSN:

2587-0335