Biopolym. Cell. 2021; 37(6):419-427.
Structure and Function of Biopolymers
Donor variability of the Wharton's jelly-derived MSCs in response to oxidative stress
1, 2Kovalchuk M. V., 2Shuvalova N. S., 1, 2Kordium V. A.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143
  2. State Institute of Genetic and Regenerative Medicine, NAMS of Ukraine
    67, Vyshhorodska Str., Kyiv, Ukraine, 04114


Aim. Since cell survival is crucial for the therapeutic efficacy, it is important to assess the MSC inter-donor variability under oxidative conditions. This study was undertaken to examine the functional responses to the oxidative stress of Wharton jelly-derived MSCs (WJ‑MSCs) from different donors. Methods. MSCs were obtained by the explant method and cultured accor-ding to standard methods. The oxidative stress was induced by hydrogen peroxide (H2O2). Using the MTT assay the treated WJ‑MSCs were analyzed for metabolic activity and survival. Results. Our findings indicated that viability of individual WJ‑MSCs was affected by H2O2 in a biphasic mode at concentrations between 6.25 and 440 µM for most cell samples. At lower H2O2concentrations, 6.25–50 μM, an increase in the viability to 16 % was noted. The maximum stimulating effect was observed at concentrations of 12.5 μM and 25 μM depending on the donor. We found that the preconditioning of WJ‑MSCs with 12.5 and 25 μM H2O2 for 24 h enhanced their survival under toxic H2O2-doses and survival rates varied for different donors. Conclusions. The findings indicated that the WJ‑MSC responses to H2O2-induced oxidative stress varied for different donors. The viability of H2O2-preconditioned WJ‑MSCs changed in a donor-dependent manner in severe environment. Consequently, to produce effective medical cells it is important to consider the donor variability in the process of oxidative MSC conditioning.
Keywords: MSCs, donor variability, oxidative stress, hormesis


[1] Tauffenberger A, Magistretti PJ. Reactive oxygen species: beyond their reactive behavior. Neurochem Res. 2021;46(1):77-87.
[2] D Li, Y Xu, CY Gao and YP Zhai. Adaptive protection against damage of preconditioning human umbilical cord-derived mesenchymal stem cells with hydrogen peroxide. Gen Mol Res. 2014;13 (3): 7304-17.
[3] Shields HJ, Traa A, Van Raamsdonk JM. Beneficial and detrimental effects of reactive oxygen species on lifespan: a comprehensive review of comparative and experimental studies. Front Cell Dev Biol. 2021; 9:628157.
[4] Calabrese EJ. Preconditioning is hormesis part I: Documentation, dose-response features and mechanistic foundations. Pharmacol Res. 2016; 110: 242-64.
[5] Gopi IK, Rattan SIS. Biphasic dose-response and hormetic effects of stress hormone hydrocortisone on telomerase-immortalized human bone marrow stem cells in vitro. Dose-Response. 2019; 17(4).
[6] Bashiri H, Amiri F, Hosseini A, Hamidi M, Roushandeh AM, Kuwahara Y, Jalili MA, Roudkena MH. Dual Pre-conditioning: a novel strategy to withstand mesenchymal stem cells against harsh microenvironments. Adv Pharm Bull. 2018; 8(3):465-70.
[7] Zhang J, Chen GH, Wang YW, Zhao J, Duan HF, Liao LM, Zhang XZ, Chen YD, Chen H. Hydrogen peroxide preconditioning enhances the therapeutic efficacy of Wharton's Jelly mesenchymal stem cells after myocardial infarction. Chin Med J (Engl). 2012;125(19):3472-8.
[8] Nouri F, Nematollahi-Mahani SN, Sharifi AM. Preconditioning of mesenchymal stem cells with non-toxic concentration of hydrogen peroxide against oxidative stress-induced cell death: the role of hypoxia-inducible factor-1. Adv Pharm Bull. 2019;9(1):76-83.
[9] Mahmoudi T, Abdolmohammadi K, Bashiri H, Mohammadi M, Rezaie MJ, Fathi F, Fakhari S, Rezaee MA, Jalili A, Rahmani MR, Tayebi L. Hydrogen peroxide preconditioning promotes protective effects of umbilical cord vein mesenchymal stem cells in experimental pulmonary fibrosis. Adv Pharm Bull. 2020; 10(1):72-80.
[10] Kang I, Lee BC, Choi SW, Lee JY, Kim JJ, Kim BE, Kim DH, Lee SE, Shin N, Seo Y, Kim H-S, Kim D-I, Kang K-S. Donor-dependent variation of human umbilical cord blood mesenchymal stem cells in response to hypoxic preconditioning and amelioration of limb ischemia. Exp Mol Med. 2018;50(4):35.
[11] Gray A, Schloss RS, Yarmush M. Donor variability among anti-infl amatory pro-activated mesenchymal stromal cells. Technology. 2016; 4 (3):201-15.
[12] Paladino FV, Sardinha LR, Piccinato CA, Goldberg AC. Intrinsic Variability present in wharton's jelly mesenchymal stem cells and T cell responses may impact cell therapy. Stem Cells Int. 2017:8492797.
[13] Trivedi A, Miyazawa B, Gibb S, Valanoski K, Vivona L, Lin M, Potter D, Stone M, Norris PJ, Murphy J, Smith S, Schreiber M, Pati S. Bone marrow donor selection and characterization of MSCs is critical for pre-clinical and clinical cell dose production J Transl Med.2019; 17:128
[14] Salehinejad P, Alitheen NB, Ali AM, Omar AR, Mohit M, Janzamin E, et al. Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton's jelly. In Vitro Cell Dev Biol Animal. 2012; 48(2):75-83.
[15] Van de Loosdrecht AA, Beelen RH, Ossenkoppele GJ, Broekhoven MG, Langenhuijsen MM. A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukaemia. J Immunol. Methods. 1994; 174 (1-2):311-320.
[16] Calabrese EJ. Hormesis and adult adipose-derived stem cells. Pharmacol Res. 2021;172:105803.
[17] Soon Yong Park, Ae-Jin Jeong, Geun-Young Kim, Ara Jo, Joo Eon Lee, Sun-Hee Leem, Joung-Hahn Yoon, Sang Kyu Ye, Jin Woong Chung. Lactoferrin Protects Human Mesenchymal Stem Cells from OxidativeStress-Induced Senescence and Apoptosis. J Microbiol Biotechnol. 2017;27(10): 1877-84.
[18] Calabrese EJ. Preconditioning is hormesis part II: How the conditioning dose mediates protection: Dose optimization within temporal and mechanistic frameworks. Pharmacol Res. 2016;110:265-75.
[19] Stavely R, Nurgali K. The emerging antioxidant paradigm of mesenchymal stem cell therapy. Stem Cells Transl Med. 2020; 9(9):985-1006.
[20] Braekke K, Harsem NK, Staff AC. Oxidative stress and antioxidant status in fetal circulation in preeclampsia. Pediat Res. 2006; 60 (5):560-564.
[21] Zhang C, Zhou L, Wang Z et al. Eradication of specific donor-dependent variations of mesenchymal stem cells in immunomodulation to enhance therapeutic values. Cell Death Dis. 2021; 12:357
[22] Boyt DT, Boland LK, Burand Jr AJ, Brown AJ, Ankrum JA. Dose and duration of interferon γ pre-licensing interact with donor characteristics to influence the expression and function of indoleamine-2,3-dioxygenase in me-senchymal stromal cells. J R Soc Interface. 2020; 17: 20190815.