Title:
METHOD FOR IN VITRO REPROGRAMMING OF FIBROBLASTS INTO SERTOLI CELLS AND APPLICATION THEREOF
Document Type and Number:
WIPO Patent Application WO/2020/006675
Kind Code:
A1
Abstract:
Disclosed is a method for in vitro reprogramming of fibroblasts into Sertoli cells and an application thereof. The method comprises the following steps: introducing a fluorescent protein reporter system specifically expressed in Sertoli cells into fibroblasts; followed by introducing a substance that increases the expression level and/or activity of the NR5A1 protein; adding G418 and culturing for 3-7 d; discarding the liquid phase, adding a medium for culturing fibroblasts and continuing culturing for 3-7 d; and separating Sertoli cells from the culture system. Experiments showed that cells prepared by the method fully acquire the characteristics of Sertoli cells, such as attracting endothelial cells, forming lipid droplets, interacting with germ cells, inhibiting lymphocyte growth and interleukin production, and having immune privilege. The invention has application value in the fields of infertility treatment, allogeneic transplantation, and cell therapy.
Inventors:
KEE KEHKOOI (CN)
LIANG JIANLIN (CN)
LIANG JIANLIN (CN)
Application Number:
PCT/CN2018/094194
Publication Date:
January 09, 2020
Filing Date:
July 03, 2018
Export Citation:
Assignee:
UNIV TSINGHUA (CN)
International Classes:
C12N5/10; C12N5/071; C12N15/65; C12N15/867
Domestic Patent References:
| WO2015147830A1 | 2015-10-01 |
Foreign References:
| CN106636210A | 2017-05-10 |
Other References:
BUGANIM, Y. ET AL.: "Direct Reprogramming of Fibroblasts into Embryonic Sertoli-Like Cells by Defined Factors", CELL STEM CELL, vol. 11, no. 3, 7 September 2012 (2012-09-07), pages 373 - 386, XP055669247
DATABASE NUCLEOTIDE 29 April 2017 (2017-04-29), PARK, M. ET AL.: "Homo Sapiens AMH 5' Regulatory Region (LOC108783649) on Chromosome 19", XP055669249, retrieved from NCBI Database accession no. NG_051647
KATO, T. ET AL.: "NR5Al is Required for Functional Maturation of Sertoli Cells during Postnatal Development", REPRODUCTION, vol. 143, no. 5, May 2012 (2012-05-01) - 31 December 2012 (2012-12-31), pages 663 - 672, XP055669252
LIANG, JIANGLIN ET AL.: "Induction of Sertoli Cells from Human Fibroblasts by NR5A1 and GATA4", BIORXIV, 29 September 2018 (2018-09-29), pages 1 - 38, XP055669260
SHIMA, Y. ET AL.: "Contribution of Leydig and Sertoli Cells to Testosterone Production in Mouse Fetal Testes", MOLECULAR ENDOCRINOLOGY, vol. 27, no. 1, 2 November 2012 (2012-11-02) - 1 January 2013 (2013-01-01), pages 63 - 73, XP055669261
See also references of EP 3805375A4
DATABASE NUCLEOTIDE 29 April 2017 (2017-04-29), PARK, M. ET AL.: "Homo Sapiens AMH 5' Regulatory Region (LOC108783649) on Chromosome 19", XP055669249, retrieved from NCBI Database accession no. NG_051647
KATO, T. ET AL.: "NR5Al is Required for Functional Maturation of Sertoli Cells during Postnatal Development", REPRODUCTION, vol. 143, no. 5, May 2012 (2012-05-01) - 31 December 2012 (2012-12-31), pages 663 - 672, XP055669252
LIANG, JIANGLIN ET AL.: "Induction of Sertoli Cells from Human Fibroblasts by NR5A1 and GATA4", BIORXIV, 29 September 2018 (2018-09-29), pages 1 - 38, XP055669260
SHIMA, Y. ET AL.: "Contribution of Leydig and Sertoli Cells to Testosterone Production in Mouse Fetal Testes", MOLECULAR ENDOCRINOLOGY, vol. 27, no. 1, 2 November 2012 (2012-11-02) - 1 January 2013 (2013-01-01), pages 63 - 73, XP055669261
See also references of EP 3805375A4
Attorney, Agent or Firm:
JEEKAI & PARTNERS (CN)
Download PDF: