Fertility and Sterility
Volume 93, Issue 7 , Pages 2385-2395, 1 May 2010

Reconstruction of endometrium in vitro via rabbit uterine endometrial cells expanded by sex steroid

  • Hai-Bin Wang, Ph.D.

      Affiliations

    • Department of Tissue Engineering, Beijing Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing, People's Republic of China
    • ,
  • Shuang-Hong Lü, Ph.D.

      Affiliations

    • Laboratory of Oncology, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, People's Republic of China
    • ,
  • Qiu-Xia Lin, Ph.D.

      Affiliations

    • Department of Tissue Engineering, Beijing Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing, People's Republic of China
    • ,
  • Li-Xin Feng, Ph.D.

      Affiliations

    • Institute of Medical Sciences, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China
    • ,
  • De-Xue Li, M.Sc.

      Affiliations

    • Department of Tissue Engineering, Beijing Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing, People's Republic of China
    • ,
  • Cui-Mi Duan, B.Sc.

      Affiliations

    • Department of Tissue Engineering, Beijing Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing, People's Republic of China
    • ,
  • Ya-li Li, M.D., Ph.D.

      Affiliations

    • Department of Obstetrics and Gynecology, General Hospital of PLA, Beijing, People's Republic of China
    • Corresponding Author InformationYa-li Li, M.D., Ph.D., Department of Obstetrics and Gynecology, General Hospital of PLA, Beijing 100853, People's Republic of China (FAX: 8610-66938044).
    • ,
  • Chang-Yong Wang, M.D., Ph.D.

      Affiliations

    • Department of Tissue Engineering, Beijing Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing, People's Republic of China
    • Corresponding Author InformationReprint requests: Chang-Yong Wang, M.D., Ph.D., Department of Tissue Engineering, Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, 27 Taiping Rd., Beijing 100850, China (FAX: 8610-68166874)

Received 12 November 2008; received in revised form 31 December 2008; accepted 14 January 2009. published online 27 March 2009.

Objective

To culture rabbit endometrial cells by using sex steroids to provide adequate seeding cells for endometrium reconstruction and uterine tissue engineering.

Design

Prospective experimental study.

Setting

Beijing Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences.

Animal(s)

New Zealand rabbit and Kunming white strain mice.

Intervention(s)

Rabbits were primed with pregnant mare serum gonadotropin and hCG. Endometrial cells were cultured with E2 and P4 of different concentrations. The endometrium was reconstructed by using endometrial cells as seeding cells and collagen–basement membrane matrix as scaffolds.

Main Outcome Measure(s)

Assay with 93-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide, immunofluorescence staining, flow cytometric analysis, hematoxylin and eosin and immunohistochemical staining, and developmental rate of embryos.

Result(s)

The expression patterns of estrogen receptor and P receptor of rabbit endometrium were different before and after treatment with pregnant mare serum gonadotropin–hCG. One hundred nanomolar E2 with 10 nmol/L P4 facilitated the proliferation of epithelial cells whereas 100 nmol/L P4 facilitated that of stromal cells. The epithelial cells could be stable if cultured for seven or eight passages. Cells in the epithelial layer of the reconstructed endometrium were cytokeratin positive. Some showed columnar morphology akin to the luminal epithelium in vivo. Reconstructed endometrium could improve the developmental rate and quality of one-cell mice embryos.

Conclusion(s)

Rabbit endometrial cells could be cultured with a long-standing proliferation capability by sex steroids and applied in uterine tissue engineering. Reconstructed endometrium with proliferated endometrial cells was akin to native endometrium in structure and function.

Key Words: Cell culture, endometrium, hCG, luteinizing hormone, collagen, tissue engineering

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 H.W. has nothing to disclose. S.L. has nothing to disclose. Q.L. has nothing to disclose. L.F. has nothing to disclose. D.L. has nothing to disclose. C.D. has nothing to disclose. Y.L. has nothing to disclose. C.W. has nothing to disclose.

 H.W. and S.L. contributed equally to this work.

 Supported by National High Technology Research and Development Program of China (No. 2006AA02A105), Nature Science Foundation of China (No. 30530220), Beijing Nature Science Foundation of China (No. 7062053), and Medical and Healthy Foundation of Military (No. 06MA318).

PII: S0015-0282(09)00141-1

doi:10.1016/j.fertnstert.2009.01.091

Fertility and Sterility
Volume 93, Issue 7 , Pages 2385-2395, 1 May 2010

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