PMID- 9822986
OWN - NLM
STAT- MEDLINE
DCOM- 19990202
LR  - 20190515
IS  - 1058-0468 (Print)
IS  - 1573-7330 (Electronic)
IS  - 1058-0468 (Linking)
VI  - 15
IP  - 9
DP  - 1998 Oct
TI  - Separating X-bearing human spermatozoa through a discontinuous Percoll density 
      gradient proved to be inefficient by double-label fluorescent in situ 
      hybridization.
PG  - 565-9
AB  - PURPOSE: Double-label fluorescence in situ hybridization (FISH) was used to 
      evaluate the efficiency of separating X- and Y-chromosome-bearing spermatozoa 
      through 12-step discontinuous Percoll gradients. METHODS: Liquefied normal semen 
      samples from 10 healthy donors were overlaid onto 25% Percoll and centrifuged. 
      Parts of the sperm pellet were saved as control, while the remaining portion was 
      separated by 12-step Percoll gradient. After centrifugation, the spermatozoa in 
      the 80% Percoll layer were collected. The X:Y ratio of the control and separated 
      spermatozoa was verified by double-label FISH (CEP SOX/SGY probes) and scored 
      blindly by one observer. Differences in the X:Y ratios between matched groups 
      were analyzed by paired t tests. RESULTS: The overall average labeling efficiency 
      was 99.2%. A significant enrichment (P = 0.02) of X-bearing spermatozoa was 
      obtained in Percoll separated fractions (mean X:Y ratio = 52.2:46.4) compared 
      with the control group (X:Y ratio = 49.5:48.4). Discontinuous Percoll gradients 
      also decreased the proportion of aneuploid spermatozoa (from 1.0 to 0.8%), but 
      the differences were nonsignificant. CONCLUSIONS: Discontinuous Percoll 
      separation did increase the X:Y ratio significantly, but the enrichment of 
      X-bearing spermatozoa is insufficient for clinical use in preconceptional sex 
      selection.
FAU - Lin, S P
AU  - Lin SP
AD  - Department of Medical Research, Mackay Memorial Hospital, Tamshui, Taipei, 
      Taiwan.
FAU - Lee, R K
AU  - Lee RK
FAU - Tsai, Y J
AU  - Tsai YJ
FAU - Hwu, Y M
AU  - Hwu YM
FAU - Lin, M H
AU  - Lin MH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Assist Reprod Genet
JT  - Journal of assisted reproduction and genetics
JID - 9206495
RN  - 0 (Chromatin)
RN  - 0 (Colloids)
RN  - 0 (DNA Probes)
RN  - 65455-52-9 (Percoll)
RN  - 7631-86-9 (Silicon Dioxide)
RN  - FZ989GH94E (Povidone)
SB  - IM
MH  - Centrifugation, Density Gradient/standards
MH  - Chromatin/chemistry
MH  - Colloids/chemistry
MH  - DNA Probes/chemistry
MH  - Female
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - Microscopy, Fluorescence
MH  - Povidone/chemistry
MH  - Sex Preselection/*methods
MH  - Silicon Dioxide/chemistry
MH  - Spermatozoa/*physiology
MH  - X Chromosome/*chemistry
MH  - Y Chromosome/*chemistry
PMC - PMC3454920
EDAT- 1998/11/21 00:00
MHDA- 1998/11/21 00:01
PMCR- 1999/10/01
CRDT- 1998/11/21 00:00
PHST- 1998/11/21 00:00 [pubmed]
PHST- 1998/11/21 00:01 [medline]
PHST- 1998/11/21 00:00 [entrez]
PHST- 1999/10/01 00:00 [pmc-release]
AID - 421706 [pii]
AID - 10.1023/a:1022590321986 [doi]
PST - ppublish
SO  - J Assist Reprod Genet. 1998 Oct;15(9):565-9. doi: 10.1023/a:1022590321986.