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

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 �li�i Mezankimal H�crelerinin HL-60 H�crelerindeki Gran�lositik Farkl�la�mas� �zerine Etkisi

Ama�: Mezankimal k�k h�creler (MKH) bir�ok h�cre tipine g�re farkl�la�abilen multipotent stromal h�crelerdir. Hematopoez s�recini d�zenleyici sitokinler ve b�y�me fakt�rleri sal�n�m� ile ve h�creler aras� etkile�im i�in �nemli h�cresel adezyon molek�llerinin ifadesi yoluyla kontrol ederler. Bu �al��ma da kemik ili�i (K�) kaynakl� MKH HL-60 h�crelerinin farkl�la�mas� �zerine etkisini morfolojik de�erlendirme, ak�m sitometri ve gen ifade analizi y�ntemleriyle ara�t�r�lmas� ama�lanm��t�r.
Gere� ve Y�ntem: K�-MKH %10 fetal s��r serumu (FSS) i�eren Dulbecco�nun modifiye Eagle ortam�nda k�lt�r edildi. ��nc� pasaj sonras�, K�-MKH 30 Gy ile ��nland�. HL-60 h�crelerinin farkl� �artlarda nas�l farkl�la�t��n� kar��la�t�rmak i�in HL-60 h�creler %10 FSS eklenmi� RPMI-1640 ortam�nda k�lt�r edildi. HL-60 h�creleri alt� kuyucuklu plaklarda all-trans retinoik asit (ATRA), K�-MKH ve ATRA ile birlikte K�-MKH ile muamele edilirken, bir kuyucu�a sadece HL60 h�creleri kondu. HL-60 h�crelerinde gran�losit alt gruplar�na �zg� genlerin ifade d�zeyleri ger�ek zamanl� polimeraz zincir reaksiyonu ile de�erlendirildi.
Bulgular: Sonu�lar�m�z K�-MKH�nin insan promiyelositik l�semi h�cre dizisi HL-60��n gran�lositik farkl�la�mas�n� destekledi�ini g�sterdi.
Sonu�: Bu �al��man�n bulgular�na g�re, K�-MKH�nin ATRA yan etkilerini azalt�c� ve hatta �nleyici etkili bir kaynak oldu�u ve k�sa ila� kullan�m� s�re�lerinde farkl�la�may� uyarabilece�i sonucunu ��kartt�k. K�-MKH etkili bir kaynak olsa da, klinik olgularda bu farkl�la�ma mekanizmas�n� iyile�tirmek i�in destekleyici ek �al��malara ihtiya� vard�r.

Anahtar Kelimeler: Mezankimal k�k h�creler, HL-60 h�creleri, Farkl�la�ma, 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

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