İnsülin, transferin ve selenöz asit preparatı kondrosit hücre sayısında artışa yol açar mı?

Year 2014, Volume: 48 Issue: 3, 313 - 319, 30.05.2014

Alper Gokce İbrahim Yilmaz Nevzat Selim Gokay Levent Can Cigdem Gokce

https://doi.org/10.3944/AOTT.2014.2635

Abstract

Amaç:Bu çalışmanın amacı insülin, insan transferin, selenöz asit (ITS) preparatının kondrosit çoğalması ve morfolojisi üzerine olumlu etkisi olduğu varsayımını test etmek ve bu eklenen maddelerin farklı hücre kültürü ortamlarındaki etkilerini biyokimyasal ve histolojik olarak araştırmak idi.

Çalışma planı: Gonartroz tanısı koyulan 57 yaşındaki bayan hastanın kıkırdak dokusundan insan kaynaklı kıkırdak hücreleri (human cartilage-derived cell, hCDC) ayrıştırıldı. Bu doku örnekleri DMEM (Dulbecco’s modified Eagle’s medium) ve RPMI-1640 içerisinde kültüre alındı. Hücrelerin kondrojenik aktiviteleri gözlendi. Hücreler, yapılan pasajlamalar sonrası, 6. hafta sonunda dört gruba ayrıldılar. 14. gün sonunda sayıları artan hücrelerin x4, x10, x20 ve x40 büyütmede mikro fotoğrafları çekildi ve invert mikroskop altında incelendiler. Yaşayabilen hücre sayısı 1. ve 14. günde MTS-ELISA hücre proliferasyonu testi ile belirlendi.

Bulgular: ITS premiks ilave edilmeyen DMEM ve ITS premiks ilave edilen RPMI-1640 kültür ortamlarında kondrojenik hücrelerin sayısının arttığı olduğu görüldü. Bu çalışmada kondrositlerin canlılığı ve sayı artışının sadece DMEM içeren Grup 3’te daha fazla olduğu gözlemlendi.

Çıkarımlar: Kültür ortamı içeriklerinin osteokondral dokudan elde edilen hücre kültürlerindeki kondrosit hücre sayısının artmasında önemli rol oynadığını düşünüyoruz.

Keywords

DMEM; hücre kültürü; ITS premiks solüsyon; kondrojenik farklılaşma; RPMI-1640.

Does insulin, transferrin and selenous acid preparation effect chondrocyte proliferation?

Abstract

Objective:The aim of this study was to test the hypothesis that insulin, human transferrin, and selenous acid (ITS) preparation have positive effects on chondrocyte proliferation and morphology and investigate the biochemical and histological effects of these additive substances in different cell culture media.

Methods: Human cartilage-derived cells (hCDCs) were isolated from the cartilage tissue of a 57-year-old woman diagnosed with gonarthrosis. Tissue samples were cultured in Dulbecco’s modified Eagle’s medium (DMEM) and RPMI-1640. The cells’ chondrogenic activities were observed. After serial passagings, cells were divided into 4 groups at the end of the 6th week. On the 14th day, proliferated cells were examined using an inverted microscope with x4, x10, x20 and x40 magnification and microphotographs were taken. Living cell quantity was determined on the first and 14th days using MTS-ELISA cell proliferation assay.

Results: DMEM (without adding ITS premix solution) and RPMI-1640 containing ITS premix solution provide proliferation of the chondrogenic cells. The proliferation and viability of chondrocytes were revealed in this study in the 3rd group (DMEM solution without additives).

Conclusion: It is suggested that the culture medium ingredients play crucial roles on chondrogenic proliferation in osteochondral tissue cultures.

Keywords

Cell culture, chondrogenic differentiation, DMEM, ITS premix solution, RPMI-1640

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