On the Cryopreservation of Individual Cells in Volumes Less than Nano Liter

B. Galmidi, Y. Shafran, R. Orvieto, N. Zurgil, M. Deutsch

2014

Abstract

About a million cases of low sperm count related diseases, are treated in one year at IVF labs, using unsuitable procedures and means for cryopreservation of individual spermatozoa. The micro-arrayed donut-shaped chamber (DSC) is a novel device for individual cell cryo-preservation which was developed and produced in the frame of the current work. Each chamber is designed to act as an individual isolated compartment, and to enclose a minimal number of sperm cells. To date, we have promising results with sperm cells that have undergone the freezing-thawing cycle in suspension volume of about 10nl-2 μl. Under the same freezing-thawing procedure, the motility of thawed sperm cells in a 1nl volume or less was unsatisfactory. The series of experiments lead us to suspect that a unique physical mechanism might be the cause for the explored phenomena. Therefore, an analytical-physical research is under way, to understand the microscopic changes in the freezing-thawing process of water, solution and cell suspension (with different geometric shapes), at very small volumes, together with a biology research aimed to understand the effect of these changes on spermatozoa. These will be followed by medical research for the development of applied clinical devices and protocols. We strongly believe that overcoming the abovementioned temporary problems of cryopreservation of individual sperm cells within volumes less than nano liter, using our novel device for cryopreservation, will ensure there is no loss of spermatozoa during the freezing-thawing process.

References

  1. A. Cesana, P. N. (2003). Sperm cryopreservation in oligoasthenospermic patients in Proceedings of spermatozoa in yolk-filled human zonae pellucidae. 75(no. 4).
  2. A. Herrler, S. E. (2006). Cryopreservation of spermatozoa in alginic acid capsules. 85(no. 1).
  3. A. Just, I. G. (2004). Novel method for the cryopreservation of testicular sperm and ejaculated spermatozoa from patients with severe oligospermia: a pilot study. Fertility and Sterility, 445-447.
  4. Adamson, M. G. (2001). A method of successful cryopreservation of small numbers of human spermatozoa. 76.
  5. C. Quintans, M. D. (2003). Development of a novel approach for cryopreservation of very small numbers of spermatozoa. 15.
  6. Cohn, j. R. (2012). Gibbs Thomson Equation . book on Demand.
  7. D. A. Isaev, S. Y. (2007). Artificial microcontainers for cryopreservation of solitary spermatozoa.
  8. DeWitt, B. L. (2007). Fundamentals of Heat and Mass Transfer. John Wiley & Sons.
  9. E. Isachenko, V. I. (2004). DNA integrity and motility of human spermatozoa after standard slow freezing versus cryoprotectant-free vitrification. 19(no. 4).
  10. E. Sereni, M. A. (2008). Freezing spermatozoa obtained by testicular fine needle aspiration: a new technique. 16(no. 1).
  11. F. Nawroth, V. I. (2002). Vitrification of human spermatozoa without cryoprotectants. 23(no. 2).
  12. Haas, F. G. (1936). A Commentary on the Scientific Writings of J. Willard Gibbs. New Haven, Connecticut: Yale University Press, 544.
  13. I. Koscinski, C. W.-K. (2007). Optimal management of extreme oligozoospermia by an appropriate cryopreservation programme. 22(no. 10).
  14. J. Liu, X. Z. (2000). Cryopreservation of a small number of fresh human testicular spermatozoa and testicular spermatozoa cultured in vitro for 3 days in an empty zona pellucida. 21(no. 3).
  15. J. O. Sohn, S. H. (2003). Comparison of recovery and viability of sperm in ICSI pipette after ultra rapid freezing or slow freezing. 80.
  16. M. Gil-Salom, J. R. (2000). Intracytoplasmic sperm injection with cryopreserved testicular spermatozoa. 169(no. 1-2).
  17. M. Montag, K. R. (1999). Laser-assisted cryopreservation of single human spermatozoa in cell- free zona pellucida. 31(no. 1).
  18. N. Bouamama, P. B. (2003). Comparison of two methods of cryoconservation of sperm when in very small numbers. 31(no. 2).
  19. N. Desai, C. C. (2004). Cryopreservation of single sperm from epidydimal and testicular samples on cryoloops: preliminary case report. 82.
  20. N. Desai, D. G. (1998). A convenient technique for cryopreservation of micro quantities of sperm. 70.
  21. N. N. Desai, H. B. (2004). Single sperm cryopreservation on cryoloops: an alternative to hamster zona for freezing individual spermatozoa. 9(no. 1).
  22. P. E. Levi-Setti, E. A. (2003). Cryopreservation of a small number of spermatozoa in yolk-filled human zonae pellucidae. 75(no. 4).
  23. Pan D, L. L. (2011). Melting the ice: on the relation between melting temperature and size for nanoscale ice crystals. ACS Nano, 4562-9.
  24. T. G. Schuster, L. M. (2003). Ultra-rapid freezing of very low numbers of sperm using cryoloops. 18(no. 4).
  25. V. Isachenko, E. I. (2005). Clean technique for cryoprotectant-free vitrification of human spermatozoa. 10(no. 3).
  26. Y. Y. Hsieh, H. D. (2000). Cryopreservation of human spermatozoa within human or mouse empty zona pellucidae. 73(no. 4).
Download


Paper Citation


in Harvard Style

Galmidi B., Shafran Y., Orvieto R., Zurgil N. and Deutsch M. (2014). On the Cryopreservation of Individual Cells in Volumes Less than Nano Liter . In Doctoral Consortium - DCBIOSTEC, (BIOSTEC 2014) ISBN Not Available, pages 88-93


in Bibtex Style

@conference{dcbiostec14,
author={B. Galmidi and Y. Shafran and R. Orvieto and N. Zurgil and M. Deutsch},
title={On the Cryopreservation of Individual Cells in Volumes Less than Nano Liter},
booktitle={Doctoral Consortium - DCBIOSTEC, (BIOSTEC 2014)},
year={2014},
pages={88-93},
publisher={SciTePress},
organization={INSTICC},
doi={},
isbn={Not Available},
}


in EndNote Style

TY - CONF
JO - Doctoral Consortium - DCBIOSTEC, (BIOSTEC 2014)
TI - On the Cryopreservation of Individual Cells in Volumes Less than Nano Liter
SN - Not Available
AU - Galmidi B.
AU - Shafran Y.
AU - Orvieto R.
AU - Zurgil N.
AU - Deutsch M.
PY - 2014
SP - 88
EP - 93
DO -