Synthesis of immunomagnetic nanoparticles and their application in the
separation and purification of CD34+hematopoietic stem cells
Wei Chen a ,Hebai Shen a ,*,Xingyu Li a ,Nengqin Jia a ,Jianming Xu b
a
Life and Environment Science College,Shanghai Normal University,Shanghai 200234,PR China
b
Zhongshan Hospital of FuDan University,Shanghai 200030,PR China
Received 31October 2005;accepted 6March 2006
Available online 18April 2006
Abstract
The silica-coated superparamagnetic nanoparticles with the uniform diameter of about 60nm were synthesized by reverse microemulsions method.And the magnetic nanoparticles were modified with N -(2-aminoethyl)-3-aminopropyltrimethoxysilane (AEAPS).The immunomagnetic nanoparticles were then successfully prepared by covalently immobilizing anti-CD34+monoclonal antibodies to the surface of amino silane modified magnetic particles.The cell separation results showed that the synthesized immunomagnetic nanoparticles could rapidly and conveniently separate the CD34+cells with high efficiency and specificity than normal ones.The surface morphology of separated target cells was examined by scanning electron microscope (SEM).Atomic force microscope (AFM)also characterized the magnetic materials on the surface of the separated target cells for the first time,which further confirmed that the target cells were separated by the immunomagnetic nanoparticles.The viability of the separated cells was studied by culturing method and Beckman Vi-cell viability analyst.Therefore,our experiments provided a new,direct,rapid mode to separate target cells.#2006Elsevier B.V .All rights reserved.
PACS:75.50.-y;07.79.LH;87.17.E
Keywords:Immunomagnetic separation;CD34+;Monoclonal antibody (MAbs);Atomic force microscopy (AFM);Scanning electron microscopy (SEM)
1.Introduction
Nanoscience is one of the most important research and development frontiers in modern science [1].The use of nanoparticle materials offers many advantages due to their unique size,physical properties,large surface area/volume and so on [2,3].Among all these materials,the magnetic particles are paid more attention.As their special trait of magnetism,they have been widely applied in various fields of bioseparation and medicine,such as protein and enzyme immobilization [4,5],immunoassay,RNA and DNA purification,cell isolation and recognition [6–8],target drug [9–11]and the PCR reaction [12].Using magnetic particles is advantageous for full automation,resulting in minimizing manual labor and providing more precise results [13].Use of nanomagnetic particles also has the advantages in assay sensitivity,rapidity and precision [14].All these bioseparation and medicine applications require that these nanoparticles have high magnetization and small diameter
with narrow particle size distribution so that the particles have uniform physical and chemical properties [15].
It is well known that the hematopoietic stem cells (HSC)are the source of all blood cells [16].They are very important for generating all kinds of cells and maintaining the number of cells in peripheric blood.Since the first successful treatment of Fanconi’s anemia by the using of human umbilical cord blood in 1988,more and more researches have been done about the transplanting of human umbilical cord blood [17].Human umbilical cord blood transplanting has been applied in curing many malignancy hematonosis,such as acute lymphocytic leukaemia,chronic granulocyte leukaemia,Hodgkin’s disease and so on.Now,it has been revealed that the human umbilical cord blood is rich in HSC.In order to study the HSC,the first thing that should be done is to separate the HSC from hematopoietic tissue.The normal way is utilizing the marker protein on the surface of HSC.Nowadays,CD34+is the most widely used marker protein of HSC.Among all those methods,the immunomagnetic separation is a very attractive method [7].Immunomagnetic separation can separate the target cells from the crude cells directly,conveniently and rapidly.As early
/locate/apsusc
Applied Surface Science 253(2006)1762–1769
*Corresponding author.Tel.:+862164322516;fax:+862164321800.E-mail address:shenhb@ (H.Shen).
0169-4332/$–see front matter #2006Elsevier B.V .All rights reserved.doi:10.1016/j.apsusc.2006.03.012
