Mesenchymal Stem Cells from Human Amniotic Membrane and Umbilical Cord Can Diminish Immunological Response in an in vitro Allograft Model

Abstract

Background/Aim: Mesenchymal stem cells (MSCs) are gaining rising interest in gynecology and obstetrics. MSCs immunomodulatory properties are suitable enough to reduce perinatal morbidity caused by inflammation in premature neonates. The aim of this study was to evaluate and compare the ability to inhibit allo-activated lymphocytes proliferation by MSCs derived from different sources: amniotic membrane (AM), umbilical cord (UC) and adipose tissue (AT). Methods: MSCs were isolated from AM (n = 7) and UC (n = 6) and AT (n = 6) of healthy women. Cells were characterized by flow cytometry and differentiation assay. To evaluate the potential of fetal and adult MSCs to diminish immunological response, mixed lymphocytes reaction (MLR) was performed. Results: Amnion and UC-derived cells displayed typical MSCs characteristics. Addition of MSCs to MLR significantly inhibited the proliferation of stimulated lymphocytes. The effect was observed regardless of the MSCs type used (p < 0.01 in all groups). Comparative analysis revealed no significant differences in this action between tested MSCs types. Additionally, no type of MSCs significantly stimulated allogeneic lymphocytes. Conclusion: The results prove the immunosuppressive capacities of fetal-derived MSCs in vitro. In the future, they may be potentially used to treat premature newborn as well as in immunomodulation in post-transplant therapy.

Comparison of the paracrine activity of mesenchymal stem cells derived from human umbilical cord, amniotic membrane and adipose tissue

“…The leading role of TGF‐β among other cytokines secreted by MSCs was established in an experiment showing that the suppression of CD4+ and CD8+ T‐cell proliferation after MSC treatment is reversed by the addition of monoclonal anti‐TGF‐β1 neutralizing antibodies . Our previous study confirmed that fetal‐derived MSCs have strong potential to diminish immunological response in mixed leukocyte reaction in vitro …”

“…However, the current leading approach is that paracrine action by MSC‐derived soluble factors is a primary mechanism by which this population enhances regeneration of the injured tissues and organs . Moreover, MSCs evoke very small to minimal immune reactivity as they suppress the proliferation of lymphocytes and are believed to be safe for allogeneic transplantation …”

“…UC samples, after removal of blood and debris, were put in sterile phosphate buffered saline (PBS) + 1% penicillin and streptomycin solution, and transported to the cell culture laboratory within 1 h. UC‐MSC isolation and culture were performed under laminar flow hood in sterile conditions. UC fragments were cut into explants (of about 3 mm 3 ) and left on a Ø 100 mm culture dish in growth medium (GM) consisting of Dulbecco's modified Eagle's Medium with low glucose (DMEM‐LG) supplemented with fetal calf serum (FCS; 10%) and antibiotic–antimycotic solution (penicillin‐streptomycin‐amphotericin; 1.5%), in standard culture conditions (37°C and 5% CO 2 atmosphere), similar to our previous experiments …”

“…The amnion was mechanically separated from the chorionic membrane. In order to eliminate potential microbiological contamination, cell isolation protocol started with immersion in 70% ethanol solution for approximately 3 s. As described in our previous experiment, samples were incubated in 0.25% trypsin–ethylenediaminetetraacetic acid solution for 60 min in 37°C while being vigorously shaken, washed twice with PBS and exposed to 1 mg/mL collagenase type I/0.025 mg/mL DNAse solution . The amnion was kept incubated in 37°C until complete dissolution of the membrane (next 60–90 min).…”

“…antibiotic-antimycotic solution (penicillinstreptomycin-amphotericin; 1.5%), in standard culture conditions (37°C and 5% CO2 atmosphere), similar to our previous experiments. 17 Isolation and culture of AM-MSCs Fragments of amniotic membranes (10 cm 2 ) were taken during the same procedure, as described above. The amnion was mechanically separated from the chorionic membrane.…”