Methods of Granulocyte Isolation from Human Blood and Labeling with Multimodal Superparamagnetic Iron Oxide Nanoparticles

Author(s)

Alvieri, Fernando ^[1] ; Mamani, Javier B. ^[1] ; Nucci, Mariana P. ^[2] ; Oliveira, Fernando A. ^[1] ; Filgueiras, Igor S. ^[1] ; Rego, Gabriel N. A. ^[1] ; de Barboza, Marycel F. ^[1] ; da Silva, Helio R. ^[1] ; Gamarra, Lionel F. ^[1]

Affiliation

^[1] Hosp Israelita Albert Einstein, BR-05652900 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Fac Med, Hosp Clin HCFMUSP, LIM44, BR-01246903 Sao Paulo - Brazil

• Document type: Journal article
• Source: Molecules; v. 25, n. 4 FEB 2 2020.
• Web of Science Citations: 0

Abstract

This in vitro study aimed to find the best method of granulocyte isolation for subsequent labeling with multimodal nanoparticles (magnetic and fluorescent properties) to enable detection by optical and magnetic resonance imaging (MRI) techniques. The granulocytes were obtained from venous blood samples from 12 healthy volunteers. To achieve high purity and yield, four different methods of granulocyte isolation were evaluated. The isolated granulocytes were labeled with multimodal superparamagnetic iron oxide nanoparticles (M-SPIONs) coated with dextran, and the iron load was evaluated qualitatively and quantitatively by MRI, near-infrared fluorescence (NIRF) and inductively coupled plasma mass spectrometry (ICP-MS). The best method of granulocyte isolation was Percoll with Ficoll, which showed 95.92% purity and 94% viability. After labeling with M-SPIONs, the granulocytes showed 98.0% purity with a yield of 3.5 x 10(6) cells/mL and more than 98.6% viability. The iron-loading value in the labeled granulocytes, as obtained by MRI, was 6.40 +/- 0.18 pg/cell. Similar values were found with the ICP-MS and NIRF imaging techniques. Therefore, our study shows that it is possible to isolate granulocytes with high purity and yield and labeling with M-SPIONs provides a high internalized iron load and low toxicity to cells. Therefore, these M-SPION-labeled granulocytes could be a promising candidate for future use in inflammation/infection detection by optical and MRI techniques. (AU)

• FAPESP's process: 16/21470-3 - Therapeutic action of mesenchymal stem cells from human bone marrow labeled with multimodal nanoparticles in diabetic rats subjected to stroke: study of cellular, molecular and functional mechanisms.
• Grantee: Lionel Fernel Gamarra Contreras
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/50983-3 - INCT 2014: complex fluids
• Grantee: Antonio Martins Figueiredo Neto
• Support Opportunities: Research Projects - Thematic Grants