A new simple and reliable vitrification device based on Hollow Fiber Vitrification (HFV) method evaluated using IVP bovine embryos

Abstract

A new vitrification device based on hollow fiber vitrification (HFV) was constructed using a glass capillary, which lead to simplified construction process and increased practicality of the device. The hollow fiber was attached to heat-pulled tip of the glass capillary using forceps. A protective sheath fitted on the capillary provided protection for the cellulose triacetate hollow fiber with loaded embryos and allowed safe storage in liquid nitrogen for long periods of time (2-12 month), transfer between tanks with liquid nitrogen and transportation within these tanks. No embryos were lost in the process. The device was tested using seven-dayold and eight-day-old IVP bovine blastocysts and expanded blastocysts as a model. Obtained survival (90% at 24 h post warming) and hatching rates (62% at 72 h post warming) of day 7 blastocysts and expanded blastocysts were comparable to those gained using various vitrification carriers. Vitrified embryos did not show an increase in the number of cells with damaged membrane or a decrease in total cell number per embryos in comparison to their non-vitrified counterparts. Day 7 and 8 expanded blastocysts did not differ significantly in terms of survival at 24 (97.01 vs. 97.50%) and 48 h post warming (95.52 vs. 95%), but showed significantly higher survival and hatching rates than day 7 and 8 blastocysts. These results indicated that high and repeatable survival rates can be obtained by selection of IVP bovine embryos at the developmental stage of expanded blastocyst for HFV. Further modification of the method may be required to achieve high and stable results with different developmental stages of IVP bovine embryo. The vitrification device presented in the current article may contribute to wider application of HFV method in livestock production.

Vitrification of mouse two‐cell and blastocyst stage embryos in simplified closed system using either a hemi‐straw or a hollow fiber device

“…The embryo vitrification using HFV device was performed as described by Matsunari et al (2012), with some modifications. In this study, we used different type of HFV with other published papers (Malenko et al, 2018; Matsunari et al, 2012; Uchikura et al, 2016) because that model was not available in our country. The modified HFV device was constructed from a 25‐mm piece of cellulose membrane dialyzers (inner diameter 200 μm, membrane thickness 15 μm, pore size 7 nm, FB‐150U; Nipro, Osaka, Japan).…”

“…Unfortunately, cellulose triacetate hollow fiber became fragile at low temperatures. Therefore, the hollow fiber was modified to prevent damage during storage in liquid nitrogen (Malenko et al, 2018; Matsunari et al, 2012). Long‐term storage using this type of modification was complex in terms of structure and sterilization.…”

Optimization of triacetate cellulose hollow fiber vitrification protocol for cryopreservation of bovine oocytes

“…Êðîìå òîãî, HFV ïîçâîëÿåò ñîáëþñòè MVC ïðè îäíîâðåìåííî âèòðèôèêàöèè ãðóïïû ýìáðèîíîâ [8,9].  íàøèõ ïðåäûäóùèõ èññëåäîâàíèÿõ â ïîëûõ âîëîêíàõ áûëè óñïåøíî âèòðèôèöèðîâàíû IVP ýìáðèîíû ÊÐÑ â âîçðàñòå 7-8 äíåé [10]. Ïðîöåäóðû âèòðèôèêàöèè/îòîãðåâàíèÿ áûëè îñíîâàíû íà ïðîòîêîëå, ïðåäëîaeåííîì Kuwayama et al [11], óñïåøíî ïðèìåíÿþùèìñÿ â ñî÷åòàíèè ñ íîñèòåëåì Cryotop äëÿ êðèîêîíñåðâàöèè îîöèòîâ è ýìáðèîíîâ ÷åëîâåêà [12,13].…”

“…Âèòðèôèêàöèÿ è îòîãðåâàíèÿ îîöèòîâ  êà÷åñòâå íîñèòåëÿ äëÿ âèòðèôèêàöèè îîöèòîâ èñïîëüçîâàëè òðèàöåòàòöåëëþëîçíûå ïîëûå âîëîêíà ñ âíóòðåííèì äèàìåòðîì 200 ìêì, òîëùèíîé ñòåíîê 15 ìêì è ðàçìåðîì ïîð 7 íì (Nipro, ßïîíèÿ) â ñîñòàâå ðàçðàáîòàííîãî íàìè óñòðîéñòâà [15]. Âèòðèôèêàöèþ è îòîãðåâàíèå îîöèòîâ ïðîâîäèëè ñ ïðèìåíåíèåì ðàíåå îïèñàííîãî ïðîòîêîëà [10] è åãî ìîäèôèêàöèé.  êà÷åñòâå áàçîâîé ñðåäû äëÿ ïðèãîòîâëåíèÿ ðàñòâîðîâ âèòðèôèêàöèè/îòîãðåâàíèÿ èñïîëüçîâàëè TLP-HEPES ñ 20% ôåòàëüíîé ñûâîðîòêè êðîâè ÊÐÑ (TH20).…”

“…Ðåçóëüòàòû è îáñóaeäåíèå Âèòðèôèêàöèÿ äîçðåâøèõ in vitro îîöèòîâ ÊÐÑ â òðèàöåòàòöåëëþëîçíûõ ïîëûõ âîëîêíàõ ñ èñïîëüçîâàíèåì ïðîòîêîëà, óñïåøíî ïðèìåíÿâøåãîñÿ äëÿ êðèîêîíñåðâàöèè 7è 8-äíåâíûõ áëàñòîöèñò è ðàñøèðåííûõ áëàñòîöèñò [10], ïðèâåëà ê ïîëó÷åíèþ êðàéíå íèçêèõ óðîâíåé âûaeèâàíèÿ (38,5%) è äðîáëåíèÿ (40,0%; òàáë.1, ãðóïïà 1). Äî ñòàäèè áëàñòîöèñòû ýìáðèîíû â äàííîé ãðóïïå íå ðàçâèâàëèñü.…”