, Volume 52, Issue 1, pp 1–11 | Cite as

Role of the embryology laboratory in the human embryonic stem cell line derivation process

  • José Luis Cortés
  • Fernando Cobo
  • Angela Helen Barnie
  • Purificación Catalina
  • Carmen Cabrera
  • Ana Nieto
  • Rosa Montes
  • Ángel Concha


With the introduction of regenerative medicine and cell therapy programmes by means of human embryonic stem cells (hESC), several research centres have begun projects of derivation of hESC lines. In some stem cell banks, such as the Andalusian Stem Cell Bank, the law also permits the creation of these cell lines. Therefore, the recovery of cryopreserved embryos, their culture and the subsequent derivation to hESC lines requires a suitable embryology laboratory and specialized and highly qualified staff. Moreover, new techniques, from therapeutic nuclear transfer, need this type of laboratory and staff, too. Several International Associations have drawn up some guidelines for laboratories where embryos are manipulated and they reflect the physical space, the staff and the equipment needed in these kinds of laboratories. Nevertheless, we can see that these guidelines do not distinguish between IVF laboratories and other laboratories that obtain hESC lines, so it would be convenient to make a distinction. Following these guidelines, we have tried to draw up concurrent aspects applicable to areas of embryology within stem cell banks. So, the design and the specific implementation programmes for these areas and other research centres with this area but which do not use IVF techniques is vital to develop embryonic cell lines in optimum conditions for future therapeutic applications, although maybe it is rather premature to standardize this type of research.


Cleanrooms Embryology laboratory Embryonic stem cell lines International guidelines IVF Stem cell bank 



Ms. Angela Helen Barnie for the English correction of the manuscript.


  1. Association of Clinical Embryologists (ACE) (2003) Accreditation Standards and Guidelines for IVF Laboratories. Scholar
  2. AGRBM (2005) Criteria for the recognition as an embryologist in human reproduction and continued professional education: the concept of the German Society for Human Reproductive Biology. Alpha 32:12–13Google Scholar
  3. American Association of Tissue Banks (2002) Standards for tissue banking, 10th edn. American Association of Tissue Banks, McLean, VAGoogle Scholar
  4. Andalucian Parliament Law 7/2003, October 20 (2003) about the regulation of the research with nonviable human embryos to “in vitro” fertilization. BOJA 210Google Scholar
  5. Antinori S, Selman HA, Caffa B et al (1996) Zona opening of human embryos using non-contact UV laser for assisted hatching in patients with poor prognosis of pregnancy. Hum Reprod 11:2488–2492Google Scholar
  6. American Society for Reproductive Medicine (ASRM) (2004) Reviewed guidelines for human embryology and andrology laboratories. www.asrm.orgGoogle Scholar
  7. Bongso A, Fong CY, Ng SC, Ratnam S (1994) Isolation and culture of inner cell mass cells from human blastocysts. Hum Reprod 9:2110–2117Google Scholar
  8. College of American Pathologist (CAP) (2002) Laboratory Improvement. www.cap.orgGoogle Scholar
  9. Castilla JA, Alcaide A, Bonada M, Expósito A, Luna C, Núňez R, de los Santos MJ (eds) (2006) I. Guidelines about Human and Physical Requirements in the Assisted Reproduction Laboratory. ASEBIR Clinical Embryology Notebooks, Madrid, SpainGoogle Scholar
  10. Cobo F, Stacey GN, Hunt C, Cabrera C, Nieto A, Montes R, Cortes JL, Catalina P, Barnie A, Concha A (2005) Microbiological control in stem cell banks: approaches to standarization. Appl Microbiol Biotechnol 68:456–466CrossRefGoogle Scholar
  11. Cobo F, Cabrera C, Catalina P, Concha A (2006a) General safety guidances in stem cell bank installations. Cytotherapy 8:47–56CrossRefGoogle Scholar
  12. Cobo F, Stacey GN, Cortes JL, Concha A (2006b) Enviromental monitoring in stem cell banks. Appl Microbiol Biotechnol 70:651–662CrossRefGoogle Scholar
  13. Cohen J, Gilligan A, Garrisi J (2004) Setting up an ART laboratory. In: Gardner DK, Weissman A, Howles CM et al (eds) Textbook of assisted reproduction techniques. Laboratory and clinical perspectives, 2nd edn. Taylor & Francis, London, UK, pp 17–24Google Scholar
  14. Commission Directive 2006/17/EC of 8 February (2006) implementing Directive 2004/23/EC of the European Parliament and of the Council as regards certain technical requirements for the donation, procurement and testing of human tissues and cells. Official Journal of the European Union, L 38/40Google Scholar
  15. Cowan CA, Klimanskaya I, McMahon J, Atienza J, Witmyer J, Zucker JP, Wang S, Morton CC, McMahon AP, Powers D, Melton DA (2004) Derivation of embryonic stem-cell lines from human blastocysts. N Engl J Med 350:1353–1356CrossRefGoogle Scholar
  16. Department of Health (2001) A code of practice for tissue banks. Department of Health, London, UKGoogle Scholar
  17. EC Guide to good Manufacturing Practice (2003) Revision to Annex 1. Manufacture of Sterile Medicinal Products. European Commission. Enterprise Directorate-GeneralGoogle Scholar
  18. European Society of Human Reproduction and Embryology (ESHRE) (2000) Guidelines for good practice in IVF laboratories. www.eshre.comGoogle Scholar
  19. European Union (2004) Directive 2004/23/EC of the European Parliament and of the Council of 31 March (2004) on setting standards of quality and safety for the donation, procurement, testing, processing, preservation, storage, and distribution of human tissues and cells. Official Journal of the European Union, L 102/48Google Scholar
  20. FSA. The Fertility Society of Australia Reproductive Technology Accreditation Committee (RTAC) (2002) Code of practice for centres using assisted reproductive technology. Scholar
  21. Hoffman LM, Carpenter MK (2005) Characterization and culture of human embryonic stem cells. Nat Biotechnol 23:699–708CrossRefGoogle Scholar
  22. ICMR-DBT. Indian Council of Medical Research (2006) Guidelines for stem cell research and therapy. www.icmr.nic.inGoogle Scholar
  23. ISO (1999) International Standard ISO 14644. Cleanrooms and associated controlled environments. Part 1: Classification of air cleanliness. International Organization for Standarization, Geneve, SwitzerlandGoogle Scholar
  24. ISO (2000) International Standard ISO 9001. Quality Management Systems-Requeriments. International Organization for Standarization, Geneve, SwitzerlandGoogle Scholar
  25. ISO (2005) International Standard ISO 17025. General requirements for the competence of testing and calibration laboratories. International Organization for Standarization, Geneve, SwitzerlandGoogle Scholar
  26. Kim HS, Oh SK, Park YB, Ahn HJ, Sung KC, Kang MJ, Lee LA, Suh CS, Kim SH, Kim DW, Moon SY (2005) Methods for derivation of human embryonic stem cells. Stem Cells 23:1228–1233CrossRefGoogle Scholar
  27. Ludwig TE, Levenstein ME, Jones JM, Berggren WT, Mitchen ER, Frane JL, Crandall LJ, Daigh CA, Conard KR, Piekarczyk MS, Llanas RA, Thomson JA (2006) Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol 24:185–187CrossRefGoogle Scholar
  28. Mortimer D (2005) A critical assessment of the impact of the European Union Tissues and Cells Directive (2004) on laboratory practices in assisted conception. Reprod Biomed Online 11:162–176Google Scholar
  29. National Academy of Medical Sciences of India (NAMSI) (2005) National Guidelines for accreditation, supervision and regulation of ART clinics in India. www.icmr.nic.inGoogle Scholar
  30. National Academies Press (NAP) (2005) Guidelines for Human Stem Cell Research. Scholar
  31. National Institute of Health (NIH) (2002) Guidance for investigators and institutional review boards regarding research involving human embryonic stem cells, germ cells, and stem cell-derived test articlesGoogle Scholar
  32. Noda Y, Goto Y, Umaoka Y et al (1994) Culture of human embryos in alpha modification of Eagle´s medium under low oxygen tension and low illumination. Fertil Steril 62:1022–1027Google Scholar
  33. Pharmaceutical Inspection Co-operation Scheme (2002) Recommendation on sterility testing. Pharmaceutical Inspection Convention, PI 012-1, November 1, 2002Google Scholar
  34. Reubinoff BE, Pera MF, Fong CY, Trounson A, Bongso A (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat Biotechnol 18:399–404CrossRefGoogle Scholar
  35. Richards M, Tan S, Fong CY, Biswas A, Chan WK, Bongso A (2003) Comparative evaluation of various human feeders for prolongued undifferentiated growth of human embryonic stem cells. Stem Cells 21:546–556CrossRefGoogle Scholar
  36. Sharp J (2005) GMP and quality assurance in sterile products manufacture. In: J Sharp (ed) Good pharmaceutical manufacturing practice. CRC Press, Florida, USA, pp 371–404Google Scholar
  37. Solter D, Knowles BB (1975) Immunosurgery of mouse blastocysts. Proc Natl Acad Sci USA 72:5099–5102CrossRefGoogle Scholar
  38. Spanish Law 14/2006, May 26th, about Assisted Reproduction Techniques 2006. BOE 126Google Scholar
  39. Spanish Royal Decree 2132\2004, about the requirements and procedures to request the development of research projects with human embryonic stem cells. 2004. BOE 262Google Scholar
  40. Tanaka N, Takeuchi T, Neri QV, Sills ES, Palermo GD (2006) Laser-assisted blastocyst dissection and subsequent cultivation of embryonic stem cells in a serum/cell free culture system: applications and preliminary results in a murine model. J Transl Med 8(4):20 DOI 10.1186/1479-5876-4-20Google Scholar
  41. Xu C, Inokuma MS, Denham J, Golds K, Kundu P, Gold JD, Carpenter MK (2001) Feeder-free growth of undifferentiated human embryonic stem cells. Nat Biotechnol 19:971–974CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • José Luis Cortés
    • 1
  • Fernando Cobo
    • 1
  • Angela Helen Barnie
    • 1
  • Purificación Catalina
    • 1
  • Carmen Cabrera
    • 1
  • Ana Nieto
    • 1
  • Rosa Montes
    • 1
  • Ángel Concha
    • 1
  1. 1.Andalusian Stem Cell Bank (Spanish Central Node)Hospital Universitario Virgen de las NievesGranadaSpain

Personalised recommendations