Induction of Umbilical Cord Blood Mesenchymal Stem Cells into Neuron-Like Cells In Vitro

  • Lingling Hou
  • Hua Cao
  • Dongmei Wang
  • Guorong Wei
  • Cixian Bai
  • Yong Zhang
  • Xuetao Pei
Case Report

Abstract

Mesenchymal stem cells (MSCs) in human umbilical cord blood are multipotent stem cells that differ from hematopoietic stem cells. They can differentiate in vitro into mesenchymal cells such as osteoblasts and adipocytes. However, differentiation into nonmesenchymal cells has not been demonstrated. Here, we report the isolation, purification, expansion, and differentiation of human umbilical cord blood MSCs into neurocytes in vitro. Cord blood samples were allowed to drain from the end of the cord into glass bottles with 20 U/mL preservative-free heparin. MSCs were isolated from human umbilical cord blood, purified, and expanded in Mesencult medium. Surface antigens of MSCs were analyzed by fluorescence-activated cell sorting (FACS). MSC passages 2,5, and 8 were induced to differentiate into neuron-like cells. Neurofilament (NF) and neuron-specific enolase (NSE) were detected by immunohistochemistry staining. Special Nissl bodies were observed by histochemical analysis. The results showed that 6.6 × 105 primary MSCs were expanded for 10 passages to obtain 9.9 × 108 MSCs, an increase of approximately 1.5 × 103-fold. FACS results showed that the MSCs did not express antigens CD34, CD11a, and CD11b and expressed CD29 and CD71, an expression pattern identical to that of human bone marrow-derived MSCs. Induction results indicated that approximately 70% of the cells exhibited a typical neuron-like phenotype. Immunohistochemistry staining suggested that induced MSCs of different passages expressed NF and NSE. Special Nissl bodies were obvious in the neuron-like cells. These results suggest that MSCs in human umbilical cord blood are capable of differentiating into neuron-like cells in vitro.Int J Hematol. 2003;78:256-261.

Key words

Umbilical cord blood mesenchymal stem cells In vitro expansion Committed induction Neuron-like cell 

References

  1. 1.
    Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells.Science. 1999;284:143–147.PubMedCrossRefGoogle Scholar
  2. 2.
    Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic tissues.Science. 1997;276:71–74.PubMedCrossRefGoogle Scholar
  3. 3.
    Woodbury D, Schwarz EJ, Prockop DJ, Black IP. Adult rat and human bone marrow stromal cells differentiate into neurons.J Neurosci Res. 2000;61:364–370.PubMedCrossRefGoogle Scholar
  4. 4.
    Deans RJ, Moseley AB. Mesenchymal stem cells: biology and potential clinical uses.Exp Hematol. 2000;28:875–884.PubMedCrossRefGoogle Scholar
  5. 5.
    Majumdar MK, Thiede MA, Mosca JD, Moorman M, Gerson SL. Phenotypic and functional comparison of cultures of marrow- derived mesenchymal stem cells (MSCs) and stromal cells.J Cell Physiol. 1998;176:57–66.PubMedCrossRefGoogle Scholar
  6. 6.
    Devine SM, Bartholomew AM, Mahmud N, et al. Mesenchymal stem cells are capable of homing to the bone marrow of non- human primates following systemic infusion.Exp Hematol. 2001;29:244–255.PubMedCrossRefGoogle Scholar
  7. 7.
    Erices A, Conget P, Minguell JJ. Mesenchymal progenitor cells in human umbilical cord blood.Br J Haematol. 2000;109:235–242.PubMedCrossRefGoogle Scholar
  8. 8.
    Pereira RF, O’Hara MD, Laptev AV, et al. Marrow stromal cells as a source of progenitor cells for nonhematopoietic tissues in transgenic mice with a phenotype of osteogenesis imperfecta.Proc Natl AcadSci USA. 1998;95:1142–1147.CrossRefGoogle Scholar
  9. 9.
    Azizi SA, Stokes D, Augelli B, DiGiolamo C, Prockop DJ. Engraftment and migration of human bone marrow stromal cells implanted in the brains of albino rat: similarities to astrocyte grafts.Proc Natl AcadSci USA. 1998;95:3908–3913.CrossRefGoogle Scholar
  10. 10.
    Cheng L, Qasba P, Vanguri P, Thiede MA. Human mesenchymal stem cells support megakaryocyte and pro-platelet formation from CD34+ hematopoietic progenitor cells.J Cell Physiol. 2000;184:58–69.PubMedCrossRefGoogle Scholar
  11. 11.
    Reese JS, Koc ON, Gerson SL. Human mesenchymal stem cells provide stromal support for efficient CD34+ transduction.J Hematother Stem Cell Res. 1999;8:515–523.PubMedCrossRefGoogle Scholar
  12. 12.
    Yoshimasu T, Tanaka R, Suenobu S, et al. Prompt and durable hematopoietic reconstitution by unrelated cord blood transplantation in a child with Fanconi anemia.Bone Marrow Transplant. 2001;27:767–769.PubMedCrossRefGoogle Scholar
  13. 13.
    Ohnuma K, Isoyama K, Ikuta K, et al. Cord blood transplantation from HLA-mismatched unrelated donors as a treatment for children with haematological malignancies.Br J Haematol. 2001;112:981–987.PubMedCrossRefGoogle Scholar
  14. 14.
    Ziegner UH, Ochs HD, Schanen C, et al. Unrelated umbilical cord stem cell transplantation for X-linked immunodeficiencies.J Pediatr. 2001;138:570–573.PubMedCrossRefGoogle Scholar
  15. 15.
    Filipovich AH. Unrelated cord blood transplantation for correction of genetic immunodeficiencies.J Pediatr. 2001;138:459–461.PubMedCrossRefGoogle Scholar
  16. 16.
    Ishii K, Katayama M, Hori K, Yodoi J, Nakanishi T. Effects of 2-mercaptoethanol on survival and differentiation of fetal mouse brain neurons cultured in vitro.Neurosci Lett. 1993;163:159–162.PubMedCrossRefGoogle Scholar

Copyright information

© The Japanese Society of Hematology 2003

Authors and Affiliations

  • Lingling Hou
    • 1
  • Hua Cao
    • 1
  • Dongmei Wang
    • 1
  • Guorong Wei
    • 1
  • Cixian Bai
    • 1
  • Yong Zhang
    • 2
  • Xuetao Pei
    • 1
  1. 1.Beijing Institute of Transfusion MedicineBeijingChina
  2. 2.Northwest Sci-Tech University of Agriculture and ForestryYangling ShaanxiChina

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