Vitrification Using Kitazato - Damia Castello, Embryologist IVI Spain

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Damia Castello an Embryologist at IVI Spain presented on vitrification using Kitazato at the RI Istanbul conference and hands on workshop on the 23rd and 24th January 2014.

He presented on the following:
- Cryotop vitrification method
- Vitrification and thawing kit
- Vitrification protocol oocyte
- Kitazato Published Scientific Results
- Closed cryotop

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Vitrification Using Kitazato - Damia Castello, Embryologist IVI Spain

  1. 1. Vitrification using Kitazato Damià Castelló www.dibimed.com
  2. 2. Cryotop Vitrification Method What is? It is a MVC device, consisting of a fine polypropilene strip, which allow the oocytes to be vitrified within very low amount of volume (0,1µl).
  3. 3. Vitrification Kit Thawing Kit
  4. 4. KITAZATO CRYOTOP VITRIFICATION: o o o o o o Valid for all stages of development, since Oocyte to Blastocyst. Cryotop SC allows vitrification in closed system for storage Vitrification Media is synthetic (free of albumin) to reduce risks. Threalose (endotoxin free) replaced Sucrose for more safety. Vitrification up to 16 Oocytes, PN, Embryos or Blastocysts in 20 min. Thawing up 16 Oocytes, PN, Embryos or Blastocysts in 20 min.
  5. 5. Protocol
  6. 6. Vitrification Protocol Oocyte
  7. 7. Vitrification Protocol Oocyte
  8. 8. Published Scientific Results
  9. 9. Outcome of cryotransfer of embryos developed from vitrified oocytes: double Em bryo d evelopm ent of fresh 'versus' vitrification has no impact on delivery rates ied metaph ase II oocytes after vitrif ICSI: a prospect ive rand om ized sibling-oocyte study /w-11 Cc>ixr,, P h O 0 . . n w t ('91otPl.a, P h 0 Ari' ... l,.. . l M I T . ( '. « r r&..AA r t t r't O Jo..e c ce S..:"'- Ph O . • • d ""n...._ f O . ·.w- t• V.,lu111,14 ti-,.. '1'. .b......_ h . l l . . 4 U • ·o:IW Prospective controlled study to evaluate laboratory and clinical outcomes of oocyte vitrification obtained in in vitro fertilization patients aged 30 to 39 years (h i1 )r1<h en C ..-.nc1 Pl1C>, Tho1 l<'>1- EI iull.. DS( • G1olnn V1i11 0 S ..• f>n"it<I B ShapifO l'.1 0., 11 , 1 ht, < , Lau ra Rienzi ', St e fania Ro man o, Lau ra A lbr icci, Robe r t a Maggiulli, Anton io Capalbo, Elena Baro ni, Silvia Co lam a ria, Fabio Sa p ie n z a, and Filippo Uba ldi G.C .R,..,'_ <..cn:rt k 1 11..cpro:j.Jed"'C r.rcocil"IC. Oh:.• lld 'c C ula. >la de .N.: r 0t.r lB, 11orre, ltNf 's . Vitrified-warmed blastocyst transfer cycles yield higher pregnancy and implantation rates compared with fresh blastocyst transfer cycle s -time for a new embryo transfer strategy? Andrew A. Toledo. M. . and b olt Pctcf Nagy. M.O..Pl'l.0. D R prco.l.it.•• t! 0 olcgf AloYJ I . ele,. 1 U n .e . ' ' '-'IW • C RYOTOP V ITRIFICAT ION AS COMPARED TO C O NVENT IONAL S LO W F REEZING FOR H UMA N EMBRYOS AT TH E C LEAVAGE STAG E: SURVIVAL AND O UTCOMES T K.M u.i 1..:·.J n T:i..1 Su' , , ... "'Sltt 1,Yl R u J , , H s u> Chil.g,Lc ns . "t J q wL f"( u , ) . f <u..,J€.f'w..do.JfJ "' '"'fril.oC.ll!") Vr.ll'MIK,,..W - . ! ' r - " 1,,f >- ':r : - . ' I I ( ,,...;klra'.. " J . o . t -Mi ...,. ' " " " . Da11da 1Zlm. :W 1 .D.." .l:umjl'< ZIUl'IJ . tlf.D.. 11Slwnr r1 Clio. , f.Sc..a Jw1qi<mg Zh< ;1,,-1Sc..:. "; ? mg. Rrum Chin ff f'IJ . Ph.T.. h . ff'1U';B f l urm.r1.f l.';r._ ,f? > .,.Xmf""ll f • .''1.f.> Ph.T.,11 TufJ r.> n . ,((. .. > :uw, .-f.O.. Ph.D..;. uud Guo Q iug Ton. . ,1-1 . JJf'.l J. 1111 .V. ' Outcomes of vitrified early cleavage-stage and blastocyst-stage embryos in a cryopreservation program: evaluation of 3,150 warming cycles ..... ( - C>.M .., · ,_, Wit f 'hO n,...,. P D 0 . , , . : . . " " -L f . . ..-c:..cwKc-rGt> h i I.> www.dibimed.com (• 0 ,._.C.0.""-" D Dibimed
  10. 10. Use of cryo-banked oocytes in an ovum donation programme: a prospective, random ized, controlled, cl inical . Marcos An a Cobo.trial Mesegue r, Jose Re mohi, and Ant o n io Pe llicer T a b l e I De m o gr ap hics, baseline ch ar act e r ist ics and ovarian stim ul ation of d onors . Egg-ba nk Number of subjects Age (years) o ocyte s re ce ive d Egg-bank N umber o f c rr bryo ' tr.m sferrcd 267 (90.S) 259 (89.6) Mean m mber of embryos .1 289 Fresh SI J ( 1.71 + 0 .7) 198 ( 1.72 + 0.7) Fresh 295 Table Ill C linical o utcome according t o t he type of 26.7 I 3.9 26.6 I 3.8 replaced 22.6 3.2 22.S 3.0 2 16 (74.7)* I I.I 2.9 11.8 2 1.8 N umber o f cycl with embryo ·re v itrificnclon c1 opre.scrvodon y 196 (66.7) Days of sti1 m•at1 on GnRHa (days) 1 1 8. 3.2 18.3 3. Mean "u1 nbcr of t e vltrrlicd or cryoprcs(t"Vcd embryos 592 (2.0 l 2.1) 74 3 (2.S l 2.3)' rFSH do se (IU) £ 2 on day of hCG (pg/ ml) 1 14 I 635 8 1774 I 62 1 lrrtplrtn tion r'a t.e 205 (39.9) 201 (10.9) 2879 2892 Pos tive hCG cest/ P• (ng /ml) 1.0 I 0.7 0.9 I 0.7 Mii oocytes retrieved 3286 ( I I. I I 3.2 3 185 (11.0 I 2.8) 1 (55.4) 48 1 (SH ) 44 48 (32.<4) 54 (37.5) BMI (Kg/m 2 me>in I ) SD 1172 120 1 3039 92.5 Urless o!ho,...-lsc lr d l t t d v t"'e' are m<.'ar- l SO or n (%). S' 11. body mass irdeoc . rfSH, roc.omhin>nr FSH. G'iRH>.GnRH "1,'0>L www.dibimed.com cle O ie• I pr 1 ncy r• t o /tr>nsf..n i » T win pregnancy rate lkiPS< c m. ln d b1"d v .LH ' " ' mean SO o r n (1'} . "P<. 0.CIS. Dibimed
  11. 11. Closed Cryotop
  12. 12. Strategies to avoid potential cross contamination Efficiency Closed systems for storage Safety
  13. 13. www.dibimed.com Dibimed
  14. 14. Vitrification Protocol For SC Cryotop
  15. 15. Objective • To compare the outcome of vitrified-banked oocytes using a semi-closed device (Cryotop-SC) to the standard procedure, employing the open Cryotop device. Primary outcome Survival rate Secondary outcomes. Clinical pregnancy rate Implantation rate Fertilization rate Embryo quality
  16. 16. Study design • Prospective, randomized, trial to test the efficiency of SC-device in our ovum donation program. • ClinicalTrials.gov NCT01745536 (Registered July 12th 2012). • IRB approval and complies with the Spanish Law of Assisted Reproductive Technologies (14/2006, 26th May). • 370 recipients (alpha 0.05 and power of 80% for sample size calculation).
  17. 17. Results Demographics, baseline characteristics and endometrial preparation SC Conventional 185 185 Nº Donations (Recipients ) Age (years) 159 42.1 4.3 146 41.1 4.5 0.488 BMI (Kg/m2) 24.2 4.2 26.3 4.2 0.155 Previous ovum-donation attempts 0.33 0.8 0.34 0.7 0.631 Days of endometrial preparation 15.1 4.6 16.0 5.4 0.651 Oocytes received (mean 2048 (12.8 2.9) 1787 (12.2 3.4) 0.191 Nº Randomized donors SD) Endometrial thickness (mm) 8.2 0.4 7.9 1.1 P value 0.459
  18. 18. Preliminary Results SC Conventional P value Survival rate 1787 (87.3) 1639 (91.7) <0.001 Oocytes inseminated 1787 (11.2 2.9) 1639 (11.9 3.4) 0.191 Fertilization rate (2PN) 1387 (74.6) 1220 (74.4) 0.393 T quality day-3 op embryos/inseminated oocyte 607 (32.6) 546 (33.3) 0.198 Blastocyst rate/inseminated oocyte 561/1144 (49.0) 571/1110(51.4) 0.254 196/561 (34.9) 201/571 (35.2) 0.924 125 (78.6) 118 (74.7) 0.360 T quality blastocyst op Nº of cycles with cryopreservation of surplus embryos Mean number of vitrified embryos 389 (3.1 2.1) 375 (3.2 2.3) 0.450
  19. 19. Preliminary Results Clinical outcome Nº transfers SC 151 (94.9) Conventional 134 (91.7) P value 0.261 Mean embryos transferred 2.0 0.71 2.0 0.92 0.567 Clinical pregnancy/cycle 89 (56.0) 68 (46.6) 0.102 Clinical pregnancy/transfer 89 (58.9) 68 (50.7) 0.052 Ongoing pregnancy/cycle 80 (50.3) 66 (45.2) 0.372 Ongoing pregnancy/transfer 80 (52.9) 66 (55.0) 0.264 Implantation rate 110 (40.8) 83 (39.3) 0.730
  20. 20. Conclusions  Although survival rate is slightly lower the clinical , outcome and general efficiency of the SC approach is similar to the Conventional Cryotop.  The SC device could be a good option when the use of the open system is discouraged due to legal restrictions.
  21. 21. Thank you!!

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