Optimization of macrophage isolation from the Persian sturgeon and the Caspian kutum fish: a comparative study

Abstract

The aim of this research was a comparative study on the isolation and culture of head kidney macrophages derived from Acipenser persicous and Rutilus frisii kutum as teleost and chondrostei species of fish. The macrophages were isolated by density gradient sedimentation, followed by adherence to a plastic surface. They exhibited strong phagocytic activity against bacteria. The effect of cell density, incubation time, FBS percentage, pH and temperatures on the cell number and viability were determined and compared. Also, the effect of light/dark regimen on viability, adherence, release of reactive oxygen species (ROS) and nitric oxide (NO) in the macrophages was determined. The results showed that the Caspian kutum macrophages were more sensitive to FBS percentage and cell density whereas the Persian sturgeon macrophages were more sensitive to pH of the cell culture media. The adherence and viability of the macrophages from both fish species firstly increased (P \ 0.05) after exposure to a light/dark regimen, but then significantly decreased as did ROS and NO productions. For the first time, this study has determined the optimal conditions for primary culture of macrophages derived from sturgeons, and shows the unique effect of light on the biology of fish immune cells.

Comprehensive update on inventory of finfish cell lines developed during the last decade (2010–2020)

“…Further, one macrophage cell line was developed from red common carp to improve Percoll gradient macrophage isolation technique and two macrophage cell lines, one from Persian sturgeon and another from Caspian kutum fish was developed to optimize macrophage isolation techniques (Qiu et al ., 2016; Rastgar et al . 2018) In this review, we point out that there was a reduction in the fish embryonic stem cell line development. Besides, there was also a reduction in fish melanoma cell culture development despite its high applicability as a model for human malignancies.…”

“…Besides, two macrophage cell lines were developed from thymus of catla and rohu to study different macrophage specific enzymes and cytokines (Chaudhary et al 2014;Rebello et al 2014). Further, one macrophage cell line was developed from red common carp to improve Percoll gradient macrophage isolation technique and two macrophage cell lines, one from Persian sturgeon and another from Caspian kutum fish was developed to optimize macrophage isolation techniques (Qiu et al, 2016;Rastgar et al 2018) In this review, we point out that there was a reduction in the fish embryonic stem cell line development.…”

“…Additionally, majority of the cell lines developed in the last decade was to isolate viruses which caused a decrease in macrophage cell culture development. However, recently, Rastgar et al (2018) standardized culture conditions of lymphocyte and macrophage cultures of Persian sturgeon and Caspian kutum fish. The seeding density was standardized to be 1 9 10 6 cells well À1 and pH 7.2-7.5.…”

“…However, recently, Rastgar et al . (2018) standardized culture conditions of lymphocyte and macrophage cultures of Persian sturgeon and Caspian kutum fish. The seeding density was standardized to be 1 × 10 6 cells well −1 and pH 7.2–7.5.…”

Potential Applications of Fluorescence-Activated Cell Sorting (FACS) and Droplet-Based Microfluidics in Promoting the Discovery of Specific Antibodies for Characterizations of Fish Immune Cells

“…erythrocytes and granulocytes) pellet to the bottom whereas mononuclear cells (i.e. macrophages/monocytes and lymphocytes) settle in an interphase layer (6)(7)(8)(9)(10)(11). Afterwards, an added separation process can be applied to further purify cells of interest.…”