ISSN: 1300-7777 E-ISSN: 1308-5263
The Effect of Bone Marrow Mesenchymal Stem Cells on the Granulocytic Differentiation of HL-60 Cells [Turk J Hematol]
Turk J Hematol. 2018; 35(1): 42-48 | DOI: 10.4274/tjh.2016.0498  

The Effect of Bone Marrow Mesenchymal Stem Cells on the Granulocytic Differentiation of HL-60 Cells

Hossein Nikkhah1, Elham Safarzadeh2, Karim Shamsasenjan1, Mehdi Yousefi2, Parisa Lotfinejad3, Mehdi Talebi1, Mozhde Mohammadian4, Farhoud Golafshan5, Ali Akbar Movassaghpour1
1Tabriz University Faculty of Medicine, Hematology and Oncology Research Center, Tabriz, Iran
2Tabriz University Faculty of Medicine, Drug Applied Research Center, Tabriz, Iran; Tabriz University Faculty of Medicine, Department of Immunology, Tabriz, Iran
3Tabriz University Faculty of Medicine, Hematology and Oncology Research Center, Tabriz, Iran; Tabriz University Faculty of Medicine, Department of Immunology, Tabriz, Iran
4Mazandaran University Faculty of Medicine, Amol Faculty of Paramedical Sciences, Sari, Iran
5Hamline University Faculty of Medicine, Department of Biology, Minnesota, USA

Objective: Mesenchymal stem cells (MSCs) are multipotent stromal cells that can differentiate into a variety of cell types. They control the process of hematopoiesis by secreting regulatory cytokines and growth factors and by the expression of important cell adhesion molecules for cell-to-cell interactions. This investigation was intended to examine the effect of bone marrow (BM)-derived MSCs on the differentiation of HL-60 cells according to morphological evaluation, flow cytometry analysis, and gene expression profile.
Materials and Methods: The BM-MSCs were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS). After the third passage, the BM-MSCs were irradiated at 30 Gy. To compare how the HL-60 cells differentiated in groups treated differently, HL-60 cells were cultured in RPMI-1640 and supplemented with 10% FBS. The HL-60 cells were seeded into six-well culture plates and treated with all-trans-retinoic acid (ATRA), BM-MSCs, or BM-MSCs in combination with ATRA, while one well remained as untreated HL-60 cells. The expression levels of the granulocyte subsetspecific genes in the HL-60 cells were assayed by real-time polymerase chain reaction.
Results: Our results revealed that BM-MSCs support the granulocytic differentiation of the human promyelocytic leukemia cell line HL-60.
Conclusion: Based on the results of this study, we concluded that BM-MSCs may be an effective resource in reducing or even preventing ATRA’s side effects and may promote differentiation for short medication periods. Though BM-MSCs are effective resources, more complementary studies are necessary to improve this differentiation mechanism in clinical cases.

Keywords: Mesenchymal stem cells, HL-60 cells, Differentiation, Alltrans-retinoic acid


Kemik İliği Mezankimal Hücrelerinin HL-60 Hücrelerindeki Granülositik Farklılaşması Üzerine Etkisi

Hossein Nikkhah1, Elham Safarzadeh2, Karim Shamsasenjan1, Mehdi Yousefi2, Parisa Lotfinejad3, Mehdi Talebi1, Mozhde Mohammadian4, Farhoud Golafshan5, Ali Akbar Movassaghpour1
1Tabriz University Faculty of Medicine, Hematology and Oncology Research Center, Tabriz, Iran
2Tabriz University Faculty of Medicine, Drug Applied Research Center, Tabriz, Iran; Tabriz University Faculty of Medicine, Department of Immunology, Tabriz, Iran
3Tabriz University Faculty of Medicine, Hematology and Oncology Research Center, Tabriz, Iran; Tabriz University Faculty of Medicine, Department of Immunology, Tabriz, Iran
4Mazandaran University Faculty of Medicine, Amol Faculty of Paramedical Sciences, Sari, Iran
5Hamline University Faculty of Medicine, Department of Biology, Minnesota, USA

Amaç: Mezankimal kök hücreler (MKH) birçok hücre tipine göre farklılaşabilen multipotent stromal hücrelerdir. Hematopoez sürecini düzenleyici sitokinler ve büyüme faktörleri salınımı ile ve hücreler arası etkileşim için önemli hücresel adezyon moleküllerinin ifadesi yoluyla kontrol ederler. Bu çalışma da kemik iliği (Kİ) kaynaklı MKH HL-60 hücrelerinin farklılaşması üzerine etkisini morfolojik değerlendirme, akım sitometri ve gen ifade analizi yöntemleriyle araştırılması amaçlanmıştır.
Gereç ve Yöntem: Kİ-MKH %10 fetal sığır serumu (FSS) içeren Dulbecco’nun modifiye Eagle ortamında kültür edildi. Üçüncü pasaj sonrası, Kİ-MKH 30 Gy ile ışınlandı. HL-60 hücrelerinin farklı şartlarda nasıl farklılaştığını karşılaştırmak için HL-60 hücreler %10 FSS eklenmiş RPMI-1640 ortamında kültür edildi. HL-60 hücreleri altı kuyucuklu plaklarda all-trans retinoik asit (ATRA), Kİ-MKH ve ATRA ile birlikte Kİ-MKH ile muamele edilirken, bir kuyucuğa sadece HL60 hücreleri kondu. HL-60 hücrelerinde granülosit alt gruplarına özgü genlerin ifade düzeyleri gerçek zamanlı polimeraz zincir reaksiyonu ile değerlendirildi.
Bulgular: Sonuçlarımız Kİ-MKH’nin insan promiyelositik lösemi hücre dizisi HL-60’ın granülositik farklılaşmasını desteklediğini gösterdi.
Sonuç: Bu çalışmanın bulgularına göre, Kİ-MKH’nin ATRA yan etkilerini azaltıcı ve hatta önleyici etkili bir kaynak olduğu ve kısa ilaç kullanımı süreçlerinde farklılaşmayı uyarabileceği sonucunu çıkarttık. Kİ-MKH etkili bir kaynak olsa da, klinik olgularda bu farklılaşma mekanizmasını iyileştirmek için destekleyici ek çalışmalara ihtiyaç vardır.

Anahtar Kelimeler: Mezankimal kök hücreler, HL-60 hücreleri, Farklılaşma, All-trans retinoik asit


Hossein Nikkhah, Elham Safarzadeh, Karim Shamsasenjan, Mehdi Yousefi, Parisa Lotfinejad, Mehdi Talebi, Mozhde Mohammadian, Farhoud Golafshan, Ali Akbar Movassaghpour. The Effect of Bone Marrow Mesenchymal Stem Cells on the Granulocytic Differentiation of HL-60 Cells. Turk J Hematol. 2018; 35(1): 42-48

Corresponding Author: Ali Akbar Movassaghpour, Iran


TOOLS
Full Text PDF
Print
Download citation
RIS
EndNote
BibTex
Medlars
Procite
Reference Manager
Share with email
Share
Send email to author

Similar articles
PubMed
Google Scholar


 



Impact Factor (2017) = 0.650