Effect of serum starvation stress on the mouse spleen mononuclear cells mixed culture: Introducing a new immunomodulatory method

Abstract

Introduction: Bone marrow and immune cell transplants are a common method of treating some blood diseases around the world. However, due to the malfunction of the immune system, its use is not always effective. In this study, we evaluated the potential of culture stress in serum-free medium in regulating the immune system. Materials and Methods: Spleen immune cells were isolated from Balb/C and C57bl/6 mice using the Ficol gradient method. C57bl/6 mice splenocytes were cultured in serum starved and non-starved conditions at indicated time points. Cell viability was determined by MTT assay. Then the allogenicity of these cells was determined by mixed lymphocytic culture (in vitro model of bone marrow transplantation) with Balb/C mouse splenocytes and cell proliferation was then examined using MTT method and necessary photographs were prepared too. Results: The viability rate of starved immune cells after 72 h was 82.1% of the control group. After 24h, the allogenicity of these cells decreased significantly compared to the control cells (P<0.01). This decrease was more pronounced at 48h and 72 h (P<0.0001). Even the response of Balb/C lymphocytes to stressed cells was less than that of background control, (P<0.0001). Morphological findings clearly confirm this reduction. Conclusion: Serum starvation induced-stress decrease allogenicity and modulates immune cells allo-reactivity. In the future, the use of mouse model could shed light on the application of this technique in bone marrow transplantation.

Keywords

• transplantation
• cell culture technique
• mouse
• immunoregulation