Display options
Cite
Handyside AH, O'Neill GT, Jones M, et al. Use of BRL-conditioned medium in combination with feeder layers to isolate a diploid embryonal stem cell line. Rouxs Arch Dev Biol. 1989;198(1):48-56doi: 10.1007/BF00376370.
Handyside, A. H., O'Neill, G. T., Jones, M., & Hooper, M. L. (1989). Use of BRL-conditioned medium in combination with feeder layers to isolate a diploid embryonal stem cell line. Roux's archives of developmental biology : the official organ of the EDBO, 198(1), 48-56. https://doi.org/10.1007/BF00376370
Handyside, Alan H, et al. "Use of BRL-conditioned medium in combination with feeder layers to isolate a diploid embryonal stem cell line." Roux's archives of developmental biology : the official organ of the EDBO vol. 198,1 (1989): 48-56. doi: https://doi.org/10.1007/BF00376370
Handyside AH, O'Neill GT, Jones M, Hooper ML. Use of BRL-conditioned medium in combination with feeder layers to isolate a diploid embryonal stem cell line. Rouxs Arch Dev Biol. 1989 May;198(1):48-56. doi: 10.1007/BF00376370. PMID: 28305783.
Copy Download .nbib Format: NLM
Share it on

Rouxs Arch Dev Biol. 1989 May;198(1):48-56. doi: 10.1007/BF00376370.

Use of BRL-conditioned medium in combination with feeder layers to isolate a diploid embryonal stem cell line.

Roux's archives of developmental biology : the official organ of the EDBO

Alan H Handyside, Gerard T O'Neill, Mary Jones, Martin L Hooper

Affiliations

  1. MRC Experimental Embryology and Teratology Unit, MRC Laboratories, Woodmansterne Road, SM5 4EF, Carshalton, Surrey, England.
  2. Department of Pathology, University of Edinburgh, Teviot Place, EH8 9AG, Edinburgh, England.
  3. Department of Anatomy, University of Edinburgh, Teviot Place, EH8 9AG, Edinburgh, England.

PMID: 28305783 DOI: 10.1007/BF00376370

Abstract

The derivation of a karyotypically normal embryonal stem (ES) cell line, E14, from inner cell masses (ICMs) isolated by immunosurgery from 129/Ola late mouse blastocysts is described. Disaggregated ICMs were cultured on mitotically-arrested fibroblast feeder layers in droplets of medium conditioned with Buffalo rat liver (BRL) cells under oil. BRL-conditioned medium inhibits the differentiation of established embryonal carcinoma (EC) and ES cell lines which can be maintained indefinitely in the complete absence of feeder cells (Smith and Hooper 1987). At clonal densities, however, a combination of BRL-conditioned medium and a feeder layer was most effective in preventing the differentiation of E14 cells. This effect was less pronounced at higher passage suggesting it may be particularly important to use a combination in the early stages of isolation. Once established, E14 has been maintained in BRL-conditioned medium alone. In non-conditioned medium on agarose, E14 cells formed embryoid bodies which when allowed to reattach differentiated into a wide variety of tissues. An HPRT-deficient sub line of E14, E14TG2a, has been demonstrated to form germline chimaeras with high efficiency after injection into blastocysts (Hooper et al. 1987). The modifications to the ES cell isolation procedure described here may improve the efficiency with which karyotypically normal lines can be derived.

Keywords: Conditioned medium; Embryonal stem cells; Feeder layer

References

  1. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3899-903 - PubMed
  2. Exp Cell Res. 1977 Feb;104(2):255-62 - PubMed
  3. Roux Arch Dev Biol. 1986 Oct;195(8):519-526 - PubMed
  4. Dev Biol. 1987 May;121(1):1-9 - PubMed
  5. Nature. 1981 Jul 9;292(5819):154-6 - PubMed
  6. Cell. 1989 Jan 27;56(2):313-21 - PubMed
  7. Chromosoma. 1973;41(2):145-58 - PubMed
  8. Proc Natl Acad Sci U S A. 1966 Jan;55(1):66-73 - PubMed
  9. J Embryol Exp Morphol. 1985 Jun;87:27-45 - PubMed
  10. Exp Cell Res. 1983 May;145(2):458-62 - PubMed
  11. Exp Cell Res. 1984 Sep;154(1):233-42 - PubMed
  12. J Embryol Exp Morphol. 1979 Aug;52:141-52 - PubMed
  13. J Reprod Fertil. 1982 Sep;66(1):161-8 - PubMed
  14. Roux Arch Dev Biol. 1987 Mar;196 (3):185-190 - PubMed
  15. Dev Biol. 1984 Jan;101(1):225-8 - PubMed
  16. Oxf Surv Eukaryot Genes. 1987;4:34-76 - PubMed
  17. J Embryol Exp Morphol. 1977 Feb;37(1):217-26 - PubMed
  18. J Reprod Fertil Suppl. 1971 Jun;14:7-21 - PubMed
  19. Nature. 1987 Mar 19-25;326(6110):292-5 - PubMed
  20. Nature. 1984 May 17-23;309(5965):255-6 - PubMed
  21. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1441-5 - PubMed
  22. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7634-8 - PubMed

Publication Types