DENTAL PULPA KAYNAKLI MEZENKİMAL KÖK HÜCRELER İÇİN ELEKTROPORASYON PARAMETRELERİNİN ARAŞTIRILMASI

Year 2016, Volume: 25 Issue: 3, 144 - 147, 01.12.2016

Zeynep Burçin Gönen

Abstract

Amaç: Bu çalışmanın amacı dental pulpa kaynaklı
mezenkimal kök hücrelere (DP-MKH) Yeşil Floresan
Protein (GFP) genini transfer etmek ve elektroporasyon
parametrelerini optimize etmektir.
Gereç ve Yöntem: DP-MKH ler GFP geni ile Neon
Transfection System kullanılarak transfekte edildi. 5
f a r k l ı e l e k t r o p o r a s y o n p a r a m e t r e s i
( 1 2 0 0 v , 2 0 m s , 1 p u l s ; 1 2 0 0 v , 2 0 m s , 2 p u l s ;
1 3 0 0 v , 4 0 m s , 1 p u l s ; 1 4 0 0 v , 2 0 m s , 1 p u l s ;
1400v,20ms,2puls) optimizasyon için karşılaştırıldı.
Transfeksiyondan sonra hücrelerin canlılık ve apoptoz
değerleri 0, 24 ve 48. saatlerde analiz edildi. Floresan
ışık yoğunluğu floresan mikroskop altında değerlendirildi.
Bulgular: DP-MKH için CD29, CD44 pozitif ve CD45
negatif bulundu. Diğer gruplara göre daha fazla canlı
hücre sayısı, GFP geninin daha yüksek ekspresyonu ve
daha düşük apoptoz 1200v, 20ms, 1puls
elektroporasyon değerinde elde edileceği tespit edildi.
Sonuç: Tavşan hayvan modeli doku mühendisliği çalışmalarında sık kullanılan bir modeldir. Çalışmamızdan
elde edilen bu sonuçlar ileri tavşan DP-MKH için optimum elektroporasyon koşullarını göstermektedir

Keywords

Yeşil floresan protein (GFP), mezenkimal stromal hücre, transfeksiyon

The Evaluation of Electroporation Parameters for Dental Pulp Mesenchymal Stem Cells

Abstract

Objective: The aim of this study was to perform Green
Fluorescent Protein (GFP) gene delivery to dental pulp
derived mesenchymal stromal cells (DP-MSC) and optimize the electroporation parameters.
Materials and Methods: DP-MSCs were transfected
with GFP gene (Neon Transfection System). Five different electroporation parameters (1200v,20ms,1puls;
1 2 0 0 v , 2 0 m s , 2 p u l s ; 1 3 0 0 v , 4 0 m s , 1 p u l s ;
1400v,20ms,1puls; 1400v,20ms,2puls) were compared
for optimization. After transfection the viability and
apoptosis of cells were analyzed in 0, 24 and 48. hours.
Fluorescent light density was examined under fluorescent microscope.
Results: CD 29 and CD 44 were positive and CD45 was
negative for DP-MSCs. Higher number of viable cells,
higher expression of GFP and less apoptosis were found
in 1200v, 20ms, 1puls electroporation parameters than
other groups in different parameters.
Conclusion: Rabbit animal model is generally used in
various tissue engineering applications. The results of
our study demonstrate optimal electroporation conditions for rabbit DP-MSCs.

Keywords

Green fluorescent protein, mesenchymal stromal cells, transfection

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