Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (36): 6776-6780.doi: 10.3969/j.issn.1673-8225.2011.36.029

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Differentiation of murine embryonic stem cells into hematopoietic stem cells induced by stromal cells in human aorta-gonad-mesonephros region and hematopoietic reconstitution in vivo

Cai Yun1, 2, Zhang Xu-chao2, 3, Chen Hui-qin1, 2, Huang Shao-liang2   

  1. 1Department of Pediatrics, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou  510630, Guangdong Province, China
    2Center for Stem Cell Research, the Second Affiliated Hospital, Sun Yat-sen University, Guangzhou  510120, Guangdong Province, China
    3Research Center of Medical Sciences, Guangdong Provincial People’s Hospital, Guangzhou  510080, Guangdong Province, China
  • Received:2011-02-19 Revised:2011-04-12 Online:2011-09-03 Published:2011-09-03
  • Contact: Zhang Xu-chao, Doctor, Associate investigator, Center for Stem Cell Research, the Second Affiliated Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China; Research Center of Medical Sciences, Guangdong Provincial People’s Hospital, Guangzhou 510080, Guangdong Province, China zhxuchao3000@yahoo.com.cn Correspondence to: Huang Shao-liang, Professor, Doctoral supervisor, Center for Stem Cell Research, the Second Affiliated Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China hshl@gzsums.edu.cn
  • About author:Cai Yun☆, Doctor, Attending physician, Department of Pediatrics, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China; Center for Stem Cell Research, the Second Affiliated Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China wcyf@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30300377*

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Abstract:

BACKGROUND: Up to now it has been confirmed that under proper conditions of hematopoietic growth factors, feeder layers of stromal cells from hematopoietic niches and their conditioned media, embryonic stem cells (ESCs) can be induced into hematopoietic stem cells (HSCs).
OBJECTIVE: To direct murine ESCs into HSCs in vitro by the support of human aorta-gonad-mesonephros (AGM) region stromal cells, and to compare the function of hematopoietic reconstitution in vivo by different routes of HSCs transplantation in mice.
METHODS: Firstly, E14 murine ESCs were induced into embryoid body (EB). Then the cells from EB were further co-cultured with human AGM region stromal cells in Transwell non-contact system for 6 days. The induced EB cells were collected for Sca-1+c-Kit+ cells analysis by flow cytometry, checked for teratoma formation and transplanted to BALB/C female mice conditioned with lethal dose 60Co γ-ray irradiation. The recipient mice were divided into four groups at random, including intravenous injection group, bilateral tibia intra-bone marrow (IBM) injection group, irradiation control group and normal control group. The survival rates, reconstitution of hematopoietic and engraftment of donor cells of different groups were monitored.
RESULTS AND CONCLUSION: Sca-1+c-Kit+ cells in EB cells after co-cultured with human AGM region stromal cells accounted for (13.12±1.30)%. Teratoma could be detected in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region stromal cells, while there was no teratoma in the mice after IBM injection of that cells. The recipients in intravenous injection group all died. The survival rate was 55.6% in IBM group, in which the peripheral blood cell count was near to the normal at day 21 after transplantation and Sry gene copies from donor could be detected. It suggested that feeder cells from human AGM region can induce directed differentiation of murine ESCs into HSCs which can reconstruct hematopoiesis in vivo without teratoma by IBM administration.

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