• The effect of animal serum replacement with Wharton's jelly on hematopoietic and mesenchymal stem cell co-culture outcomes
  • Yoda Yaghoubi,1 Majid Zamani,2 Mehdi Yousefi,3,* AliAkbar Movasaghpour,4 Amir Mehdizadeh,5 Alireza Pishgahi,6
    1. Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
    2. Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
    3. Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
    4. Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
    5. Endocrine Research Center, Tabriz university of Medical Sciences, Tabriz, Iran
    6. Physical Medicine and Rehabilitation research Center, Tabriz University of Medical Science, Iran


  • Introduction: The clinical use of stem cell transplantation over the last decade has been accepted and is a suitable and practical cell strategy for many cancers, blood malignancies and immune system disorders. One of stem cell sources is human umbilical cord blood. Compared to other sources, cord blood has fewer CD34+ hematopoietic stem cells. Therefore, in order to enhance the clinical efficacy of autologous and allogeneic hematopoietic stem cell transplantation, in vitro culture and passage of these cells is critical. The main constituents of the culture medium are animal serums. However, some of the limitations of these serums include various pathogens (prions and mycoplasmas) transmission risk, and possible immune responses against animal antigens. In addition, resources for access to animal serum are declining. One alternative is Wharton's jelly which contains a range of growth factors such as platelet-derived growth factor (PDGF), epithelial growth factor (EGF), fibroblast basic growth factor (bFGF), acidic fibroblast growth factor (a FGF), transforming growth factor beta1 (TGF-β1) and Insulin-like growth factor (IGF-I). For most of these molecules, the receptor is present on stem cells and precursors, affecting cell function. Therefore, this study aimed to evaluate the effect of animal serum replacement with Wharton's jelly on hematopoietic and mesenchymal stem cell co-culture outcomes.
  • Methods: The human umbilical cord blood was obtained from consenting parturients who delivered a full-term infants and then Mononuclear cells (MNCs) were isolated from cord blood using ficoll hypaque (density, 1077). The CD34+/MNC fraction was isolated with superparamagnetic microbeads selection using high-gradient magnetic field and mini MACS columns. Cd34+ (1×104 Cell/ml) were cultured in IMDM medium containing 20% Wharton's jelly along with SCF, FIT-3L and TPO cytokines on MSC feeder layer for 7 days. After 7 days, CD34 expression was assessed by flow cytometry. The total nucleated cells was also evaluated. CD34+ culture in FBS-containing medium was considered as control.
  • Results: After 7 days, 10% and 9% CD34+ cells population were observed after FBS and Wharton's jelly treatment following a flowcytometric assay, respectively. In addition, 8 and 7-fold expansion were observed in media containing FBS and Wharton's jelly, respectively.
  • Conclusion: Our data indicate that not only Wharton’s jelly shows similar effects to FBS in culture media, but also reduces the risk of xenoviruses transmission and immunological responses.
  • Keywords: Wharton's jelly, animal serum, hematopoietic stem cell, cell therapy, mesenchymal stem cells