Author/Authors :
Dehdilani، Nima نويسنده Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran , , Shamsasenjan، Karim نويسنده Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran. , , Movassaghpour، Aliakbar نويسنده Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. , , Akbarzadehlaleh، Parvin نويسنده Pharmaceutical Biotechnology Department, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran , , Amoughli Tabrizi، Bahram نويسنده Department of Clinical Sciences, Faculty of Veterinary Medicine, Tabriz Branch, Islamic Azad University, Tabriz, Iran , , Parsa، Hamed نويسنده Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran , , Sabagi، Fatemeh نويسنده Iran Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran ,
Abstract :
Objective: Three-dimensional (3D) biomimetic nanofiber scaffolds have widespread applications
in biomedical tissue engineering. They provide a suitable environment for cellular
adhesion, survival, proliferation and differentiation, guide new tissue formation and
development, and are one of the outstanding goals of tissue engineering. Electrospinning
has recently emerged as a leading technique for producing biomimetic scaffolds with micro
to nanoscale topography and a high porosity similar to the natural extracellular matrix
(ECM). These scaffolds are comprised of synthetic and natural polymers for tissue engineering
applications. Several kinds of cells such as human embryonic stem cells (hESCs)
and mouse ESCs (mESCs) have been cultured and differentiated on nanofiber scaffolds.
mESCs can be induced to differentiate into a particular cell lineage when cultured as embryoid
bodies (EBs) on nano-sized scaffolds.
Materials and Methods: We cultured mESCs (2500 cells/100 ?l) in 96-well plates with
knockout Dulbecco’s modified eagle medium (DMEM-KO) and Roswell Park Memorial
Institute-1640 (RPMI-1640), both supplemented with 20% ESC grade fetal bovine serum
(FBS) and essential factors in the presence of leukemia inhibitory factor (LIF). mESCs
were seeded at a density of 2500 cells/100 ?l onto electrospun polycaprolactone (PCL)
nanofibers in 96-well plates. The control group comprised mESCs grown on tissue culture
plates (TCP) at a density of 2500 cells/100 ?l. Differentiation of mESCs into mouse
hematopoietic stem cells (mHSCs) was performed by stem cell factor (SCF), interleukin-3
(IL-3), IL-6 and Fms-related tyrosine kinase ligand (Flt3-L) cytokines for both the PCL and
TCP groups. We performed an experimental study of mESCs differentiation.
Results: PCL was compared to conventional TCP for survival and differentiation of
mESCs to mHSCs. There were significantly more mESCs in the PCL group. Flowcytometric
analysis revealed differences in hematopoietic differentiation between the PCL and
TCP culture systems. There were more CD34+ (Sca1+) and CD133+ cells subpopulations
in the PCL group compared to the conventional TCP culture system.
Conclusion: The nanofiber scaffold, as an effective surface, improves survival and
differentiation of mESCs into mHSCs compared to gelatin coated TCP. More studies
are necessary to understand how the topographical features of electrospun fibers affect
cell growth and behavior. This can be achieved by designing biomimetic scaffolds
for tissue engineering.
