Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (27): 5046-5049.doi: 10.3969/j.issn.1673-8225.2010.27.024

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Ex vivo expanded human umbilical cord blood mononuclear cells and hematopoietic and immune reconstitution with cytokine combination in NOD/SCID mice

Xu Li1, Lin Jin-ying1, Mao Ping2   

  1. 1 Department of Hematology, People’s Hospital, Nanning  530021, Guangxi Zhuang Autonomous Region, China; 2 Department of Hematology, Guangzhou Municipal First People’s Hospital, Guangzhou 510180, Guangdong Province, China
  • Online:2010-07-02 Published:2010-07-02
  • About author:Xu Li★, Master, Attending physician, Department of Hematology, People’s Hospital, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:

    the Grant of Health Department of Guangxi Zhuang Autonomous Region of China, No. Z2008068*; the Key Program for Science and Technology of Science and Technology Bureau of Guangzhou City, No. 2002z2-E0241*

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Abstract:

BACKGROUND: In vitro amplification of cytokine-mediated cord blood hematopoietic cells can solve the lack of quantity during cord blood transplantation.
OBJECTIVE: To explore the effect of the proper combination cytokines without stroma on the expansion of the mononuclear cells from umbilical cord blood in vitro, and to evaluate the effect of stem cell factor + FLT3 ligand + thrombopoietin on the hematopoietic and immune reconstitution.
METHODS: Human fresh cord blood mononuclear cells were cultured in serum-free and stroma-free medium containing different combination of cytokines for 7 days. According to the different combination of cytokines, our experiment was divided into groups. Mononuclear cells from cord blood were expanded for 7 days in culture containing stem cell factor + FLT3 ligand + thrombopoietin, and the expanded cells were infused into sublethally irradiated NOD/SCID mice. At days 0 and 7, numbers of cord blood CD34+ cells and CD34+CXCR4+, CD34+CD49d+, CD34+CD62L+ cells were assayed. Six weeks after transplantation, the human cells in mice were assessed by flow cytometry and PCR.
RESULTS AND CONCLUSION: Human CD45+ cells could be detected in the recipients 6 weeks after infusion. Detection rate of human specific genes and the survival rate of NOD/SCID mice implanted with expanded cells were higher than mice transplanted with fresh sample and saline (P < 0.05). The human cells of medullary system cells (CD33+), T lymphocytes (CD4+), B lymphocytes (CD19+) and hemopoietic stem cells (CD34+) were detected in bone marrow of NOD/SCID mice. Results indicated that the ex vivo expansion of umbilical cord blood mononuclear cells using the combination of 3 cytokines (stem cell factor + FLT3 ligand + thrombopoietin) is sufficient for clinical application. The expanded cord blood mononuclear cells can insert and reconstruct hematopoietic and immune functions of NOD/SCID mice.

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