四分之一1PN會在20-24h post IVF形成2PN

16-18h post IVF觀察胚胎原核(PN), 單一原核胚胎1PN機率約 5%

4h後, 25%1PN會在20-22h post IVF形成2PN, 這些胚胎發展與正常胚胎無異

TimeLaps顯示60%胚胎之PN為一前一後asynchrony形成

1PN基因分析顯示單套染色體46%, 雙套染色體29%, mosaicism染色體25%

http://humrep.oxfordjournals.org/content/8/2/221.abstract?ijkey=ccb573e3a5f5208c360fe8c21c1ed238cf9da072&keytype2=tf_ipsecsha

1PN機轉

單一原核1PN胚胎
70%正常受孕但PN形成過程障礙或
30%孤雌生殖parthenogenetic

IVF較ICSI易形成1PN(2倍機率)

http://humrep.oxfordjournals.org/content/14/7/1869.long

hCG100-150iu誘導排卵可達不錯IVF懷孕率

IVF誘導排卵COH配合FSH+hCG100-150iu可達不錯懷孕率

http://humrep.oxfordjournals.org/content/27/10/3074.full

3PN胚胎可存活於4度達24h

3PN胚胎可存活於4度達24h

http://www.ncbi.nlm.nih.gov/pubmed/23820799

PCO濾泡會分泌較一般濾泡較高之AMH

多囊性卵巢PCO病患單一基礎濾泡會分泌較一般正常病患卵巢濾泡較高之AMH

http://www.fertstert.org/article/S0015-0282(14)02293-6/abstract

使用time-lapse 觀察胚胎分裂速度及型態可研判胚胎性別

使用胚胎即時監測系統time-lapse, 觀察胚胎分裂速度及型態可研判胚胎性別

http://www.fertstert.org/article/S0015-0282(14)02310-3/abstract

新一代自動化玻璃化冷凍胚胎與卵子儀器

Gavi 新一代自動化玻璃化冷凍儀器

Gavi - The World's First Automated Vitrification Instrument

冷凍胚胎與卵子臨床效果類似傳統cryotop玻璃化冷凍法

https://www.youtube.com/watch?v=EXcq77IGG2s

http://humrep.oxfordjournals.org/content/29/11/2431.abstract

IMSI對IVF懷孕結果無明顯助益

IMSI (傳高倍數挑選精蟲顯微注射)vs. ICSI(傳統精蟲顯微注射)

IMSI對IVF懷孕結果無明顯助益

http://www.ncbi.nlm.nih.gov/pubmed/23884963

Zona pellucida opening 優於ZP thinning

動物實驗顯示

中空透明帶Zona pellucida (ZP) opening可能優於ZP thinning

http://www.ncbi.nlm.nih.gov/pubmed/25267786

陰道施與黃體素Luteal support明顯優於口服黃體素

陰道施與黃體素安胎效果Luteal support明顯優於口服黃體素

http://humupd.oxfordjournals.org/content/13/6/581.full

IVF短療程懷孕率類似長療程

短療程short protocol懷孕率類似長療程long protocol
COH針劑費用較便宜

http://www.ncbi.nlm.nih.gov/pubmed/18492366?dopt=Abstract

P4過高易造成懷孕率下降

60000 IVF case顯示
P4過高(>正常質0.8ng/ml)易造成懷孕率下降

P4過高對於冷凍胚胎懷孕率無明顯影響

http://humupd.oxfordjournals.org/content/19/5/433.abstract

COH後,子宮內膜PR 表現上升, ER表現下降

COH後,子宮內膜PR 表現上升, ER表現下降

優質胚胎可能可克服較差(P4上升)之內膜環境

Day 2-3胚胎著床率IR約10-15%

http://humrep.oxfordjournals.org/content/20/6/1541.full

囊胚期胚胎植入可能可降低過早黃體化不良影響

過早黃體化(施打hCG當天hCG>1.5, 機率約18.2%)---可能影響胚胎著床率

延後胚胎植入至囊胚期胚胎植入可能可降低其不良影響

http://www.ncbi.nlm.nih.gov/pubmed/17555751?dopt=Abstract

蜂蜜& Vit E 有助於精蟲生長

動物實驗顯示，蜂蜜& Vit E 有助於精蟲生長

http://www.jhrsonline.org/article.asp?issn=0974-1208;year=2013;volume=6;issue=1;spage=54;epage=58;aulast=Hemadi

Embryo loading方式比較

Embryo loading方式(In Group A, the embryos were drawn directly into the ET catheter from culture microdrop under the oil. In Group B, the embryos were transferred from culture microdrop into G2 medium in center-well dish. )
比較


http://www.jhrsonline.org/article.asp?issn=0974-1208;year=2013;volume=6;issue=1;spage=65;epage=69;aulast=Halvaei

Impact of different embryo loading techniques on pregnancy rates in in vitro fertlizaton/embryo transfer cycles

Background: Embryo transfer (ET) technique is one of the important factors of in vitro fertlization success. Among the different steps in ET technique, less attention has been given to embryo loading (EL). The aim was to compare the impact of two different techniques of EL on pregnancy rate in IVF/ET cycles. Materials and Methods: In this retrospective study, 144 and 170 patients were placed in groups A and B, respectively. In Group A, the embryos were drawn directly into the ET catheter from culture microdrop under the oil. In Group B, the embryos were transferred from culture microdrop into G2 medium in center-well dish. Then, the embryos were drawn into the catheter and finally transferred into the uterus. Both groups were adjusted for other parameters based on the EL technique. The main outcome measure was pregnancy rate. Results: There were insignificant differences for etiology of infertility, source of sperm, type of stimulation protocol, percent of IVF or intracytoplasmic sperm injection type of ET catheter, cycles with good quality embryos and transferred embryos between two groups. The rate of both chemical and clinical pregnancy was higher in Group B compared to A, but the difference was insignificant (P = 0.09 and P = 0.1, respectively). Conclusion: It seems that there is no difference in the outcome by loading the embryo from microdrop or center-well dish.

Table 1: Comparison of different clinical characteristics between groups A (embryo drawn directly from microdrop) and B (embryo drawn from transfer medium)

縮減囊胚腔體積可能可提高冷凍解凍懷孕率

縮減囊胚期胚胎之囊胚腔體積，可能可提高冷凍解凍懷孕率

http://www.rbej.com/content/7/1/99

性生活對於IVF胚胎植入懷孕率並無明顯影響

植入胚胎前後性生活，對於胚胎懷孕率並無明顯影響，甚至可能可提高胚胎著床率

http://humrep.oxfordjournals.org/content/15/12/2653.full

無精症檢查

無精症病患需
測FSH, Testosterone, PRL,
尿液離心精蟲檢查，睪丸切片
PE檢查(靜脈曲張，睪丸萎縮，隱睪，副睪)
染色體分析

Low FSH, low Testosterone---原發性低性腺男性不孕症hypogonadotropic hypogonadism
Normal FSH & Testosterone---可能是阻塞性無精症 ，睪丸先天異常

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3093801/?report=reader

培養液長期放置可能對胚胎有不良影響

培養液長期放置於37度C培養箱(overnight)可能對胚胎有不良影響

prolong euilibration of culture medium might be detrimental for embryo.

http://www.ncbi.nlm.nih.gov/pubmed/15705308?dopt=Abstract&holding=f1000,f1000m,isrctn

Reprod Biomed Online. 2005 Jan;10(1):124-9.

胚葉細胞越多對於外界環境之耐受性越高

2-cell比PN時期胚胎更能忍受高溫之培養環境

胚葉細胞越多對於外界環境之耐受性越高
http://www.ncbi.nlm.nih.gov/pubmed/1627931

溫度下降易傷害細胞質內胞器

胚胎細胞核(PN) vs 胚胎細胞質

胚胎細胞質對於溫度(下降)變化之敏感性較細胞核為高

溫度下降容易造成胚胎細胞質內胞器&紡錘體傷害

http://www.ncbi.nlm.nih.gov/pubmed/9211428

低養培養對胚胎是否明顯有益仍充滿爭議

低氧培養(5%O2) vs 大氣培養(19.3%O2)
對胚胎受孕(D0)及早期發育達(D0-3)無明顯差異

對胚胎晚期(D4,5)&著床是否明顯有助益仍充滿爭議

http://www.rbej.com/content/9/1/143

IVF藥物會影響子宮內膜之基因表現

誘導排卵COH藥物會影響子宮內膜之基因表現，進而干擾胚胎著床

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871799/?report=reader

hyaluronidase denuding

hyaluronidase denuding

80iu/ml, 250um pipetting, 60s
--->改為
10iu/ml, 1000um(1mm) pipetting, 130s

Conclusion:
Denuding
<80 (maybe 40 iu/ml) hyaluronidase
<60s
250um pipetting

http://humrep.oxfordjournals.org/content/12/10/2246.long

40 iu hyaluronidase使用於cumulus removal

40 iu/ml hyaluronidase使用於cumulus removal

recombinanet hyaluronidase vs. bovine hyaluronidase (Type VIII, Sigma Chemical Company, St Louis, MO, USA) 
效果類似

http://humrep.oxfordjournals.org/content/23/8/1815.long

根據精蟲品質決定IVF時間長短

根據精蟲品質，活動力，數量，過去懷孕病史
決定IVF精蟲卵子受孕時間長短

http://www.ncbi.nlm.nih.gov/pubmed/22136561

玻璃化冷凍與慢速冷凍之比較

玻璃化冷凍與傳統慢速冷凍之比較
vitrification vs. slow freezing

玻璃化冷凍
胚胎存活率 95%
胚葉細胞存活率 75%

傳統慢速冷凍
胚胎存活率 88%
胚葉細胞存活率 45%

Modified 傳統慢速冷凍(提高sucrose濃度)
可提高胚胎存活率

https://www.youtube.com/watch?v=FHggyWPxCkE

3PN形成原因牽涉卵細胞質異常

多原核3PN形成原因，除都多重受精外，亦可能牽涉卵細胞質異常

http://www.ncbi.nlm.nih.gov/pubmed/15589875

IVF後6h未受孕卵施行rescue ICSI

IVF後6h 檢查PB以確定是否受孕
未受孕卵施行rescue ICSI, 懷孕率12%

http://www.ncbi.nlm.nih.gov/pubmed/16580372/

多餘卵子捐贈後施行ICSI-ET，懷孕率達47%

取卵超過20顆卵子可將多餘卵子冷凍並不明顯影響PR

多餘卵子捐贈後施行ICSI-ET，懷孕率達47%

http://www.ncbi.nlm.nih.gov/pubmed/12513887/

ET管應距fundus 1cm

ET管應距fundus 1cm (5-15mm)
<5mm 流產機率較高

http://www.rbej.com/content/11/1/114

Pill主要差異為黃體素之種類不同

Pill主要差異為黃體素之種類不同
Pill組成包括
雌激素 (30 μg ethinylestradio) & 
黃體素 (etroprogesterone (i.e. dydrogesterone), progesterone derivatives (i.e. medrogestone) 17α-hydroxyprogesterone derivatives (i.e. chlormadinone acetate, cyproterone acetate, medroxyprogesterone acetate, megestrol acetate), 19-norprogesterone derivatives (i.e. nomegestrol, promegestone, trimegestone, nesterone), 19-nortestosterone derivatives norethisterone (NET), lynestrenol, levonorgestrel, desogestrel, gestodene, norgestimate, dienogest) and spironolactone derivatives (i.e. drospirenone).)

http://www.sciencedirect.com/science/article/pii/S0378512208003514

培養液之白蛋白具去除毒素效果

培養液之白蛋白albumin(2 mg/ml )具減除toxin毒素效果
使用不含albumin之培養液可提高sperm motility assay之敏感性

20ml syringe對胚胎有相對毒性

http://humrep.oxfordjournals.org/content/15/7/1586.long

VitroLife 封閉式玻璃冷凍系統

VitroLife 封閉式玻璃冷凍系統

http://www.vitrolife.com/en/Fertility/Products/Vitrification/Rapid-i-Vitrification-System/

1-cell 胚胎比2-cell 胚胎對培養環境更為敏感

1-cell 胚胎(2PN)比2-cell 胚胎對培養環境更為敏感

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2842869/

IMSI偵測精蟲頭部之空泡以提高ICSI懷孕率

高倍顯微鏡(6600x)可偵測精蟲頭部之空泡，藉以挑選無空泡精蟲施行ICSI (IMSI)
以提高懷孕率

http://humrep.oxfordjournals.org/content/21/7/1787.full

卵泡成長週期中有2-3個生長波

自然狀況下人類排卵時卵泡大小約2cm,
卵泡成長週期中，約有2-3個生長波

http://www.cbra.org.br/pages/publicacoes/animalreproduction/issues/download/v6n1/pag%2020-29.pdf

卵泡生長波由多次複合生長波組成

卵泡生長波由多次複合生長波組成，包括主波, 次波, 3rd波
主波形成之優勢卵泡較次波, 3rd波形成之優勢卵泡為大

http://www.sciencedirect.com/science/article/pii/S0737080607800128

COH反應以COH 4天後卵巢反應為準

AMH, AFC是COH前預測卵巢反應之最有效工具

但COH劑量調整仍應以COH 4天後卵巢反應為準

http://humrep.oxfordjournals.org/content/25/1/221.full

濾泡每日生長速度1-2mm

濾泡生長速度約1-4mm/d (平均1.33mm/d)

濾泡中期(day9)以後, 濾泡>0.9cm才有可能成熟

http://humupd.oxfordjournals.org/content/18/1/73.full#ref-22

LH peak後24h排卵

P4 在LH peak前開始上升
LH peak後24h排卵
E2 peak 在LH peak之前

高齡病患
FSH上升,
E2略升,
AMH下降, IGF下降, inhibin下降

http://humupd.oxfordjournals.org/content/18/1/73.full#ref-22

Day 1 胚胎PN, PB發展順序

Day 1 胚胎PN, PB發展順序
2nd PB->male PN->female PN
2nd PB會在ICSI後8hr內排出(平均2.5hr)
female PN較靠近2nd PB
平均male PN(21um)略大於female PN(19um)
核仁male PN略多於female PN

PN會逐漸長大
male PN 17->24 um
female PN 15->22 um

http://humrep.oxfordjournals.org/content/12/3/532.long

ICSI未受孕主因: 卵細胞未被激活

ICSI後卵子未受孕主因: 卵細胞未被激活

http://www.ncbi.nlm.nih.gov/pubmed/9663780

囊胚有收縮現象

研究顯示囊胚期胚胎具有自我收縮現象
此現象與胚胎孵化有關

http://www.ncbi.nlm.nih.gov/pubmed/14967891

https://www.jstage.jst.go.jp/article/jrd/49/6/49_6_413/_pdf

濾泡成長經歷多濾泡成長波

濾泡成長過程可經歷多個賀爾蒙刺激形成之濾泡成長波follicular wave

http://www.biolreprod.org/content/69/3/1023.full

囊胚期植入易產生同卵雙胞胎

囊胚期植入較易產生同卵雙胞胎 (3.9%)

http://www.ncbi.nlm.nih.gov/pubmed/?term=Da+Costa%2C+Hum+Reprod+2001

2次人工授孕有助益男性不孕症

施行2次人工授孕對於精蟲品質不佳之病患有助益

http://humrep.oxfordjournals.org/content/26/3/576.full

pill對子宮內膜基因表現無明顯影響

IVF前使用pill，對子宮內膜基因表現無明顯不良影響

http://humrep.oxfordjournals.org/content/early/2014/04/03/humrep.deu065.abstract

低劑量類固醇對一般IVF病患並無明顯助益

低劑量類固醇對一般IVF病患並無明顯助益
對特定IVF病患(自體免疫, 排斥)可能有益

http://humrep.oxfordjournals.org/content/17/6/1544.full

父母B形肝炎可能經由IVF傳染至胚胎或卵子

父母B形肝炎可能經由IVF治療過程傳染至胚胎或卵子

http://humrep.oxfordjournals.org/content/26/7/1860.full

使用pill會抑制LH

COH前使用pill會抑制COH過程之LH，可能下降IVF懷孕率

http://humrep.oxfordjournals.org/content/21/2/352.full

GnRHantagonist療程不需事前使用pill

GnRHantagonist療程不需事前使用pill

使用pill 並不明顯提高懷孕率
反而提高COH劑量

http://www.ncbi.nlm.nih.gov/pubmed/18054003?dopt=Abstract

P4過高對於囊胚期植入較無明顯影響

P4>1.5 ng/ml對於囊胚期植入較無明顯影響

囊胚約百分之40具染色體異常

Day 3胚胎約50-60%具有染色體異常

Day 5胚胎約40%具有染色體異常

囊胚期胚胎最易出現異常之染色體為Chr 21,22,X,Y

http://humrep.oxfordjournals.org/content/23/11/2596.full

PGD是否異常無法全然代表未來胚胎是否正常‧

PGD目前最被質疑之處為相當高比例本身同時具有正常與不正常之胚葉細胞(mosaicism ，10-90%不正常之胚葉細胞)
57%之Day 3胚胎 & 45%之Day 5胚胎具mosaicism現象

由於胚胎具自我修復異常細胞現象，PGD所檢驗胚葉細胞是否異常無法全然代表未來胚胎是否正常‧

http://humrep.oxfordjournals.org/content/21/1/223.full