Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?

İlktan Baştan; Ergun Akçay

Review

Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?

Year 2019, Volume: 59 Issue: Ek Sayı, 130 - 136, 04.06.2020

İlktan Baştan , Ergun Akçay

Abstract

Her ülkenin, kendine özgü hayvan genetik kaynaklarını, nesli tükenme tehlikesine karşın veya gelecekte gen aktarım çalışmaları için koruması, önemli bir stratejik planlamadır. Gen bankalarında sıvı azot bağımlı olarak saklanan spermatozoon genetik materyallerinin, sıvı azot ile kontaminasyon riskinin bulunması ve sıvı azot ikmali aksaklıklarına bağlı olarak, spermatozoonların zarar görerek fertilizasyon yeteneğini kaybetmesi muhtemeldir. Son yıllarda sıvı azot ile spermatozoon saklama modeline alternatif olarak gösterilen spermatozoon liyofilizasyonu (dondurup-kurutarak saklama) işlemi sonucunda, spermatozoon motilite kaybı ve hücre membran hasarı görülmesine rağmen DNA bütünlüğü korunmaktadır. Liyofilize spermatozoon genetik materyalinin, intra-sitoplazmik spermatozoon enjeksiyonu (ICSI) ile laboratuvar hayvanları başta olmak üzere yaban hayvanları ve çiftlik hayvanlarında in vitro fertilizasyon uygulaması ile yavru alınması mümkündür. Bu derlemede liyofilize spermatozoon protokolleri, saklama koşulları ve in vitro fertilizasyon çalışmaları hakkında bilgi verilmesi amaçlanmıştır.

Keywords

Gen bankası, kriyokonservasyon, dondurarak- kurutma, yardımcı üreme teknikleri

References

1. Abdalla H, Hirabayashi M, Hochi S (2009): The ability of freeze- dried bull spermatozoa to induce calcium oscillations and resumption of meiosis. Theriogenology, 71(3): 543–52.
2. Adams GDJ, Cook I, Ward KR (2015): The principles of freze-drying. Cryopreservation and Freeze-Drying Protocols. Ed.: Wolkers WF, Oldenof H, s:121-143.
3. Akın AO (2017): Çiftlik Hayvanları genetik kaynaklarının korunması ve sürdürülebilir kullanımı küresel stratejilerinin ve Türkiye örneğinin değerlendirilmesi. Doktora Tezi, Ankara Üniversitesi Fen Bilimleri Enstitüsü.
4. Ak K, Cirit Ü, Sandal Aİ, Arıcı R, Özmen MF (2017): Dondurulmuş Spermanın Depolanması ve Kullanımında Saha Koşullarında Yapılan Hatalı Uygulamalar ve Çözüm Önerileri. Turkiye Klinikleri Reproduction and Artificial Insemination-Special Topics, 3(1): 44-59.
5. Anzalone DA, Palazzese L, Iuso D, Martino G, Loi P (2018): Freeze-dried spermatozoa: An alternative biobanking option for endangered species. Animal Reproduction Science, 190: 85–93.
6. Arav A, Saragusty J (2016): Directional freezing of sperm and associated derived technologies. Animal Reproduction Science, 169: 6–13.
7. Benedict M (1905): The determination of water in foods and physiological preparations. Arn J Physiol, 13: 309–329.
8. Bialy G, Smith VR (1957): Freeze-drying of bovine spermatozoa. Journal of Dairy Science, 40: 739-745.
9. Billy D, Potts M (2001): Life and death of dry prokaryotes. Res Microbiol, 153: 7–12.
10. Birleşmiş Milletler Gıda ve Tarım Örgütü (FAO) (2012): Croyoconservation of animal genetic resources. FAO Animal Production and Health Guidelines No. 12. Rome.
11. Choi YH, Varner D, Love CC, Hartman DL, Hinrichs K (2011): Production of live foals via intracytoplasmic injection of lyophilized sperm and sperm extract in the horse. Reproduction, 142: 529-538.
12. Derijik AA, Heijden GW, Giele M, Philippens ME, Van Bayel CC, Boer P (2006): Gamma H2AX signaling during sperm chromatin remodelling in the mouse zygote. DNA Repair 5: 959–971.
13. Ergün Z (2015). Biyolojik maddelerin kurutularak saklanması: liyofilizasyon. Etlik Vet Mikrobiyoloji Dergisi, 26(1): 35-40.
14. Flosdorf EW, Mudd S (1935): Procedure and apparatus for preservation in “lyophile” form of serum and other biological substances. The J. of Immunology, 29(5): 389-425.
15. Flosdorf EW, Kımball AC (1939): Studies with H. Pertussis: Maintenance of Cultures in Phase I. J Bacteriol 35: 696–704.
16. Gianaroli L, Magli MC, Stanghelline I, Crippa A, Crivello B, Pescatorı ES, Ferrarettı AP (2012): DNA integrity is maintained after freeze-drying of human spermatozoa. Fertility and sterility, 97(5): 1067-1073.
17. Gil L, Olaciregui M, Luño V, Malo C, González N, Martínez F (2014). Current Status of Freeze-Drying Technology to Preserve Domestic Animals Sperm. Reproduction in Domestic Animals, 49: 72–81.
18. Hara H, Tagiri M, Hwang IS, Takahashi M, Hirabayashi M, Hochi S (2014): Adverse effect of cake collapse on the functional integrity of freeze-dried bull spermatozoa. Cryobiology, 68(3): 354–360.
19. Harrouk W, Codrington A, Vinson R, Robaire B, Hales BF (2000): Paternal exposure to cyclophosphamide induces DNA damage and alters the expression of DNA repair genes in the rat preimplantation embryo. Mutation Research, 461: 229–241.
20. Hochi S, Abdalla H, Hara H, Hirabayashi M (2011): Challenging endeavour for preservation of freeze-dried mammalian spermatozoa. Journal of Reproduction and Development, 57(5): 557-563.
21. Jaroudi S, Kakourou G, Cawood S, Doshi A, Ranieri DM, Serhal P (2009): Expression profiling of DNA repair genes in human oocytes and blastocysts using microarrays. Human Reproduction, 24: 2649–2655.
22. Kaneko T, Whittingham DG, Yanagimachi R (2003a): Effect of pH value of freeze-drying solution on the chromosome integrity and developmental ability of mouse spermatozoa. Biology of reproduction, 68(1): 136-139.
23. Kaneko T, Whittingham DG, Overstreet JW, Yanagimachi R (2003b): Tolerance of the mouse sperm nuclei to freeze-drying depends on their disuliyofilizeide status. Biology of reproduction, 69(6): 1859-1862.
24. Kaneko T, Nakagata N (2006): Improvement in the long-term stability of freeze-dried mouse spermatozoa by adding of a chelating agent. Cryobiology, 53(2): 279-282.
25. Kaneko T, Serikawa T (2012): Successful long-term peservation of rat sperm by freeze-drying. Cryobiology, 64(3): 211- 214.
26. Kaneko T, Ito H, Sakamoto H, Onuma M, Inoue-Murayama M (2014): Sperm preservation by freeze-drying for the conservation of wild animals. PloS one, 9(11): e113381.
27. Kaneko T (2016): Sperm freeze-drying and micro-insemination for biobanking and maintenance of genetic diversity in mammals. Reproduction, Fertility and Development, 28(8): 1079-1087.
28. Kawase Y, Araya H, Kamada N, Jishage K, Suzuki H (2005): Possibility of long-term preservation of freeze-dried mouse spermatozoa. Biology of reproduction, 72(3): 568-573.
29. Keskintepe L, Pacholczyk G, Machnicka A, Norris K, Curuk MA, Khan I, Brackett BG (2002): Bovine blastocyst development from oocytes injected with freeze-dried spermatozoa. Biology of Reproduction, 67(2): 409–415.
30. Keskintepe L, Eroğlu A (2015): Freeze-drying of Mammalian Sperm. Cryopreservation and Freeze-Drying Protocols. Ed.: Wolkers WF, Oldenof H, p:489-497.
31. Krokida MK, Karathanos VT, Maroulis ZB (1998): Effect of freeze-drying conditions on shrinkage and porosity of dehydrated agricultural products. Journal of Food Engineering, 35(4): 369–380.
32. Kusakabe H, Yanagimachi R, Kamiguchi Y (2007): Mouse and human spermatozoa can be freeze-dried without damaging their chromosomes. Human Reproduction, 23(2): 233–239.
33. Kwon IK, Park KE, Niwa, K (2004): Activation, pronuclear formation, and development in vitro of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa. Biology of Reproduction, 71(5): 1430–1436.
34. Liu JL, Kusakabe H, Chang CC, Suzuki H, Schmidt DW, Julian M, Yang X (2004): Freeze-dried sperm fertilization leads to full-term development in rabbits. Biology of Reproduction, 70(6): 1776–1781.
35. Lopez-fernandez C, Fernandez JL, Gosalbez A, Arroyo F, Vazquez JM, Holt WV, Gosalvez J (2008): Dynamics of sperm DNA fragmentation in domestic animals: Ram. Theriogenology, 70(6): 898-908.
36. Malik A, Laily M, Zakir MI (2015): Effects of long term storage of semen in liquid nitrogen on the viability, motility and abnormality of frozen thawed Frisian Holstein bull spermatozoa. Asian Pacific Journal of Reproduction, 4(1): 22–25.
37. Men NT, Kikuchi K, Furusawa T, Dang-Nguyen TQ, Nakai M, Fukuda A, Tajima A (2016): Expression of DNA repair genes in porcine oocytes before and after fertilization by ICSI using freeze-dried sperm. Animal Science Journal, 87(11): 1325– 1333.
38. Morishita N, Ochi M, Horiuchi T (2019): Development of golden hamster embryos effectively produced by injection of sperm heads sonicated in Tris-HCl buffer with EGTA. Reproductive Medicine and Biology, 18(1): 83–90.
39. Nireeaha GR, Divya L, Sowmya C, Venkateshan N, Babu MN, Lavakumar V (2013): Lyophilization/freeze drying—an review. International Journal of Novel Trends in Pharmaceutical Sciences, 3(4): 87-98.
40. Nur Z, Birler S, Üstüner B, Zık B, Özdaş ÖB, Akkoç CÖ (2009-2012). Koç spermasının liyofilizasyonu ve intra-stoplazmik sperm enjeksiyonu yöntemi ile embriyo üretimi. (Yayınlanmamış veriler) TÜBİTAK Projesi, TEBAG 108R018.
41. Olaciregui M, Luño V, Gonzalez N, De Blas I, Gil L (2015): Freeze-dried dog sperm: Dynamics of DNA integrity. Cryobiology, 71(2): 286–290.
42. Olaciregui M, Luño V, Martí JI, Aramayona J, Gil L (2016): Freeze-dried stallion spermatozoa: evaluation of two chelating agents and comparative analysis of three sperm DNA damage assays. Andrologia, 48(9): 988–994.
43. Olaciregui M, Luño V, Domingo P, González N, Gil L (2017): In vitro developmental ability of ovine oocytes following intracytoplasmic injection with freeze-dried spermatozoa. Scientific Reports, 7(1): 1096.
44. Polge C, Smith AU, Parkes AS (1949): Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, 164(4172): 666.
45. Ringleb J, Waurich R, Wibbelt G, Streich WJ, Jewgenow K (2011): Prolonged storage of epididymal spermatozoa does not affect their capacity to fertilise in vitro-matured domestic cat (Felis catus) oocytes when using ICSI. Reproduction, Fertility and Development, 23(6): 818.
46. Shackell LF (1909): An improved method of desiccation, with some applications to biological problems. Arn J Physiol 20: 325–340.
47. Shahba MI, El-Sheshtawy RI, El-Azab AS, Abdel-Ghaffar AE, Zıada MS, Zaky AA (2016): The effect of freeze-drying media and storage temperature on ultrastructure and DNA of freeze- dried buffalo bull spermatozoa. Asian Pacific Journal of Reproduction, 5(6): 524-535.
48. Sherman JK (1954): Freezing and freeze-drying of human spermatozoa. Fertility and sterility, 5(4): 357-371.
49. Şen U (2013): Liyofilize spermanın in vitro embriyo üretiminde kullanımı. 8.Ulusal Zootekni Bilim Kongresi, 5-7 Eylül, Bildiri Kitabı, s:183-188.
50. Wakayama T, Yanagimachi R (1998): Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nature Biotechnology, 16(7): 639–641.
51. Wossidlo M, Arand J, Sebastiano V, Lepikhov K, Boiani M, Reinhardt R, Walter J (2010): Dynamic link of DNA demethylation, DNA strand breaks and repair in mouse zygotes. The EMBO journal, 29(11): 1877-1888.
52. Yanagımachi R (1994): Mammalian fertilization. The physiology of reproduction, Knobil E, Neill JD, Eds.: Raven Press, p:189-317.
53. Yavaş İ, Cantekin Z, Korkmaz Yavaş T (2013): Sıvı azot ile yapılan kryopreservasyon tekniklerinde mikrobiyal kontaminasyon riskleri ve çözüm önerileri. Lalahan Hay. Arşt. Enst. Derg. 53(1): 39-45
54. Yöney T (2005): Gıdaları dondurarak kurutup saklamak nasıl çalışır? Bilim teknik dergisi, 12:101.

Year 2019, Volume: 59 Issue: Ek Sayı, 130 - 136, 04.06.2020

İlktan Baştan , Ergun Akçay

Abstract

References

1. Abdalla H, Hirabayashi M, Hochi S (2009): The ability of freeze- dried bull spermatozoa to induce calcium oscillations and resumption of meiosis. Theriogenology, 71(3): 543–52.
2. Adams GDJ, Cook I, Ward KR (2015): The principles of freze-drying. Cryopreservation and Freeze-Drying Protocols. Ed.: Wolkers WF, Oldenof H, s:121-143.
3. Akın AO (2017): Çiftlik Hayvanları genetik kaynaklarının korunması ve sürdürülebilir kullanımı küresel stratejilerinin ve Türkiye örneğinin değerlendirilmesi. Doktora Tezi, Ankara Üniversitesi Fen Bilimleri Enstitüsü.
4. Ak K, Cirit Ü, Sandal Aİ, Arıcı R, Özmen MF (2017): Dondurulmuş Spermanın Depolanması ve Kullanımında Saha Koşullarında Yapılan Hatalı Uygulamalar ve Çözüm Önerileri. Turkiye Klinikleri Reproduction and Artificial Insemination-Special Topics, 3(1): 44-59.
5. Anzalone DA, Palazzese L, Iuso D, Martino G, Loi P (2018): Freeze-dried spermatozoa: An alternative biobanking option for endangered species. Animal Reproduction Science, 190: 85–93.
6. Arav A, Saragusty J (2016): Directional freezing of sperm and associated derived technologies. Animal Reproduction Science, 169: 6–13.
7. Benedict M (1905): The determination of water in foods and physiological preparations. Arn J Physiol, 13: 309–329.
8. Bialy G, Smith VR (1957): Freeze-drying of bovine spermatozoa. Journal of Dairy Science, 40: 739-745.
9. Billy D, Potts M (2001): Life and death of dry prokaryotes. Res Microbiol, 153: 7–12.
10. Birleşmiş Milletler Gıda ve Tarım Örgütü (FAO) (2012): Croyoconservation of animal genetic resources. FAO Animal Production and Health Guidelines No. 12. Rome.
11. Choi YH, Varner D, Love CC, Hartman DL, Hinrichs K (2011): Production of live foals via intracytoplasmic injection of lyophilized sperm and sperm extract in the horse. Reproduction, 142: 529-538.
12. Derijik AA, Heijden GW, Giele M, Philippens ME, Van Bayel CC, Boer P (2006): Gamma H2AX signaling during sperm chromatin remodelling in the mouse zygote. DNA Repair 5: 959–971.
13. Ergün Z (2015). Biyolojik maddelerin kurutularak saklanması: liyofilizasyon. Etlik Vet Mikrobiyoloji Dergisi, 26(1): 35-40.
14. Flosdorf EW, Mudd S (1935): Procedure and apparatus for preservation in “lyophile” form of serum and other biological substances. The J. of Immunology, 29(5): 389-425.
15. Flosdorf EW, Kımball AC (1939): Studies with H. Pertussis: Maintenance of Cultures in Phase I. J Bacteriol 35: 696–704.
16. Gianaroli L, Magli MC, Stanghelline I, Crippa A, Crivello B, Pescatorı ES, Ferrarettı AP (2012): DNA integrity is maintained after freeze-drying of human spermatozoa. Fertility and sterility, 97(5): 1067-1073.
17. Gil L, Olaciregui M, Luño V, Malo C, González N, Martínez F (2014). Current Status of Freeze-Drying Technology to Preserve Domestic Animals Sperm. Reproduction in Domestic Animals, 49: 72–81.
18. Hara H, Tagiri M, Hwang IS, Takahashi M, Hirabayashi M, Hochi S (2014): Adverse effect of cake collapse on the functional integrity of freeze-dried bull spermatozoa. Cryobiology, 68(3): 354–360.
19. Harrouk W, Codrington A, Vinson R, Robaire B, Hales BF (2000): Paternal exposure to cyclophosphamide induces DNA damage and alters the expression of DNA repair genes in the rat preimplantation embryo. Mutation Research, 461: 229–241.
20. Hochi S, Abdalla H, Hara H, Hirabayashi M (2011): Challenging endeavour for preservation of freeze-dried mammalian spermatozoa. Journal of Reproduction and Development, 57(5): 557-563.
21. Jaroudi S, Kakourou G, Cawood S, Doshi A, Ranieri DM, Serhal P (2009): Expression profiling of DNA repair genes in human oocytes and blastocysts using microarrays. Human Reproduction, 24: 2649–2655.
22. Kaneko T, Whittingham DG, Yanagimachi R (2003a): Effect of pH value of freeze-drying solution on the chromosome integrity and developmental ability of mouse spermatozoa. Biology of reproduction, 68(1): 136-139.
23. Kaneko T, Whittingham DG, Overstreet JW, Yanagimachi R (2003b): Tolerance of the mouse sperm nuclei to freeze-drying depends on their disuliyofilizeide status. Biology of reproduction, 69(6): 1859-1862.
24. Kaneko T, Nakagata N (2006): Improvement in the long-term stability of freeze-dried mouse spermatozoa by adding of a chelating agent. Cryobiology, 53(2): 279-282.
25. Kaneko T, Serikawa T (2012): Successful long-term peservation of rat sperm by freeze-drying. Cryobiology, 64(3): 211- 214.
26. Kaneko T, Ito H, Sakamoto H, Onuma M, Inoue-Murayama M (2014): Sperm preservation by freeze-drying for the conservation of wild animals. PloS one, 9(11): e113381.
27. Kaneko T (2016): Sperm freeze-drying and micro-insemination for biobanking and maintenance of genetic diversity in mammals. Reproduction, Fertility and Development, 28(8): 1079-1087.
28. Kawase Y, Araya H, Kamada N, Jishage K, Suzuki H (2005): Possibility of long-term preservation of freeze-dried mouse spermatozoa. Biology of reproduction, 72(3): 568-573.
29. Keskintepe L, Pacholczyk G, Machnicka A, Norris K, Curuk MA, Khan I, Brackett BG (2002): Bovine blastocyst development from oocytes injected with freeze-dried spermatozoa. Biology of Reproduction, 67(2): 409–415.
30. Keskintepe L, Eroğlu A (2015): Freeze-drying of Mammalian Sperm. Cryopreservation and Freeze-Drying Protocols. Ed.: Wolkers WF, Oldenof H, p:489-497.
31. Krokida MK, Karathanos VT, Maroulis ZB (1998): Effect of freeze-drying conditions on shrinkage and porosity of dehydrated agricultural products. Journal of Food Engineering, 35(4): 369–380.
32. Kusakabe H, Yanagimachi R, Kamiguchi Y (2007): Mouse and human spermatozoa can be freeze-dried without damaging their chromosomes. Human Reproduction, 23(2): 233–239.
33. Kwon IK, Park KE, Niwa, K (2004): Activation, pronuclear formation, and development in vitro of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa. Biology of Reproduction, 71(5): 1430–1436.
34. Liu JL, Kusakabe H, Chang CC, Suzuki H, Schmidt DW, Julian M, Yang X (2004): Freeze-dried sperm fertilization leads to full-term development in rabbits. Biology of Reproduction, 70(6): 1776–1781.
35. Lopez-fernandez C, Fernandez JL, Gosalbez A, Arroyo F, Vazquez JM, Holt WV, Gosalvez J (2008): Dynamics of sperm DNA fragmentation in domestic animals: Ram. Theriogenology, 70(6): 898-908.
36. Malik A, Laily M, Zakir MI (2015): Effects of long term storage of semen in liquid nitrogen on the viability, motility and abnormality of frozen thawed Frisian Holstein bull spermatozoa. Asian Pacific Journal of Reproduction, 4(1): 22–25.
37. Men NT, Kikuchi K, Furusawa T, Dang-Nguyen TQ, Nakai M, Fukuda A, Tajima A (2016): Expression of DNA repair genes in porcine oocytes before and after fertilization by ICSI using freeze-dried sperm. Animal Science Journal, 87(11): 1325– 1333.
38. Morishita N, Ochi M, Horiuchi T (2019): Development of golden hamster embryos effectively produced by injection of sperm heads sonicated in Tris-HCl buffer with EGTA. Reproductive Medicine and Biology, 18(1): 83–90.
39. Nireeaha GR, Divya L, Sowmya C, Venkateshan N, Babu MN, Lavakumar V (2013): Lyophilization/freeze drying—an review. International Journal of Novel Trends in Pharmaceutical Sciences, 3(4): 87-98.
40. Nur Z, Birler S, Üstüner B, Zık B, Özdaş ÖB, Akkoç CÖ (2009-2012). Koç spermasının liyofilizasyonu ve intra-stoplazmik sperm enjeksiyonu yöntemi ile embriyo üretimi. (Yayınlanmamış veriler) TÜBİTAK Projesi, TEBAG 108R018.
41. Olaciregui M, Luño V, Gonzalez N, De Blas I, Gil L (2015): Freeze-dried dog sperm: Dynamics of DNA integrity. Cryobiology, 71(2): 286–290.
42. Olaciregui M, Luño V, Martí JI, Aramayona J, Gil L (2016): Freeze-dried stallion spermatozoa: evaluation of two chelating agents and comparative analysis of three sperm DNA damage assays. Andrologia, 48(9): 988–994.
43. Olaciregui M, Luño V, Domingo P, González N, Gil L (2017): In vitro developmental ability of ovine oocytes following intracytoplasmic injection with freeze-dried spermatozoa. Scientific Reports, 7(1): 1096.
44. Polge C, Smith AU, Parkes AS (1949): Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, 164(4172): 666.
45. Ringleb J, Waurich R, Wibbelt G, Streich WJ, Jewgenow K (2011): Prolonged storage of epididymal spermatozoa does not affect their capacity to fertilise in vitro-matured domestic cat (Felis catus) oocytes when using ICSI. Reproduction, Fertility and Development, 23(6): 818.
46. Shackell LF (1909): An improved method of desiccation, with some applications to biological problems. Arn J Physiol 20: 325–340.
47. Shahba MI, El-Sheshtawy RI, El-Azab AS, Abdel-Ghaffar AE, Zıada MS, Zaky AA (2016): The effect of freeze-drying media and storage temperature on ultrastructure and DNA of freeze- dried buffalo bull spermatozoa. Asian Pacific Journal of Reproduction, 5(6): 524-535.
48. Sherman JK (1954): Freezing and freeze-drying of human spermatozoa. Fertility and sterility, 5(4): 357-371.
49. Şen U (2013): Liyofilize spermanın in vitro embriyo üretiminde kullanımı. 8.Ulusal Zootekni Bilim Kongresi, 5-7 Eylül, Bildiri Kitabı, s:183-188.
50. Wakayama T, Yanagimachi R (1998): Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nature Biotechnology, 16(7): 639–641.
51. Wossidlo M, Arand J, Sebastiano V, Lepikhov K, Boiani M, Reinhardt R, Walter J (2010): Dynamic link of DNA demethylation, DNA strand breaks and repair in mouse zygotes. The EMBO journal, 29(11): 1877-1888.
52. Yanagımachi R (1994): Mammalian fertilization. The physiology of reproduction, Knobil E, Neill JD, Eds.: Raven Press, p:189-317.
53. Yavaş İ, Cantekin Z, Korkmaz Yavaş T (2013): Sıvı azot ile yapılan kryopreservasyon tekniklerinde mikrobiyal kontaminasyon riskleri ve çözüm önerileri. Lalahan Hay. Arşt. Enst. Derg. 53(1): 39-45
54. Yöney T (2005): Gıdaları dondurarak kurutup saklamak nasıl çalışır? Bilim teknik dergisi, 12:101.

There are 54 citations in total.

Details

Primary Language	Turkish
Subjects	Zootechny (Other)
Journal Section	Collection
Authors	İlktan Baştan 0000-0001-8155-1960 Ergun Akçay This is me 0000-0002-7491-5671
Publication Date	June 4, 2020
Published in Issue	Year 2019 Volume: 59 Issue: Ek Sayı

Cite

APA	Baştan, İ., & Akçay, E. (2020). Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi, 59(Ek Sayı), 130-136.
AMA	Baştan İ, Akçay E. Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi. June 2020;59(Ek Sayı):130-136.
Chicago	Baştan, İlktan, and Ergun Akçay. “Spermatozoon Liyofilizasyonu: Hayvan Genetik kaynaklarının Korunması için Yeni Bir Saklama Modeli Olabilir Mi?”. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 59, no. Ek Sayı (June 2020): 130-36.
EndNote	Baştan İ, Akçay E (June 1, 2020) Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 59 Ek Sayı 130–136.
IEEE	İ. Baştan and E. Akçay, “Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?”, Lalahan Hayvancılık Araştırma Enstitüsü Dergisi, vol. 59, no. Ek Sayı, pp. 130–136, 2020.
ISNAD	Baştan, İlktan - Akçay, Ergun. “Spermatozoon Liyofilizasyonu: Hayvan Genetik kaynaklarının Korunması için Yeni Bir Saklama Modeli Olabilir Mi?”. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 59/Ek Sayı (June 2020), 130-136.
JAMA	Baştan İ, Akçay E. Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi. 2020;59:130–136.
MLA	Baştan, İlktan and Ergun Akçay. “Spermatozoon Liyofilizasyonu: Hayvan Genetik kaynaklarının Korunması için Yeni Bir Saklama Modeli Olabilir Mi?”. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi, vol. 59, no. Ek Sayı, 2020, pp. 130-6.
Vancouver	Baştan İ, Akçay E. Spermatozoon liyofilizasyonu: Hayvan genetik kaynaklarının korunması için yeni bir saklama modeli olabilir mi?. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi. 2020;59(Ek Sayı):130-6.

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