Korean J Fertil Steril.  2004 Mar;31(1):29-39.

Improvement of Pregnancy Rate in Preimplantation Genetic Diagnosis with FISH Procedure by the Laboratory Optimization and Experiences

• Lim CK
• Min DM
• Lee HS
• Byun HK
• Park SY
• Ryu HM
• Kim JY
• Koong MK
• Kang IS
• Jun JH

Abstract

OBJECTIVES
This study was performed to evaluate the laboratory system for successful PGD using fluorescence in situ hybridization (FISH) and the clinical outcome of PGD cycles in five years experiences.
METHODS
A total of 181 PGD-FISH cycles of 106 couples were performed, and diagnosed chromosome normality in the preimplantation embryos. The laboratory and clinical data were classified by the following optimization steps, and statistically analyzed. Phase I: Blastomere biopsy with two kinds of pipettes, removal of cytoplasmic proteins without treatment of pepsin and culture of biopsied embryos with single medium; Phase II: Blatomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with single medium; Phase III: Blastomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with sequential media.
RESULTS
A total of 3,209 oocytes were collected, and 83.8% (2,212/2,640) of fertilization rate was obtained by ICSI procedure. The successful blastomere biopsies were accomplished in 98.6% (2,043/2,071) of embryos, and the successful diagnosis rate of FISH was 94.7% (1,935/ 2,043) of blastomeres from overall data. Embryo transfers with normal embryos were conducted in 93.9% (170/181) of started cycles. There was no difference in the successful rate of biopsy and diagnosis among Phase I, II and III. However, the pregnancy rate per embryo transfer of Phase III (38.8%, 26/67) was significantly (p<0.05) higher than those of Phase I (13.9%, 5/36) and Phase II (14.9%, 10/67).
CONCLUSIONS
The laboratory optimization and experience for the PGD with FISH procedure can increase the pregnancy rate to 38.8% in the human IVF-ET program. Our facility of PGD with FISH provides the great possibility to get a normal pregnancy for the concerned couples by chromosomal aberrations.

Keyword

Preimplantation genetic diagnosis (PGD); Fluorescence in-situ hybridization (FISH); Chromosome; Pregnancy rate

MeSH Terms

Biopsy
Blastocyst
Blastomeres
Chromosome Aberrations
Cytoplasm
Diagnosis
Embryo Transfer
Embryonic Structures
Family Characteristics
Fertilization
Fluorescence
Humans
In Situ Hybridization
Oocytes
Pepsin A
Pregnancy Rate*
Pregnancy*
Preimplantation Diagnosis*
Prostaglandins D
Sperm Injections, Intracytoplasmic
Pepsin A
Prostaglandins D