40-80%Percoll無法有效率的分離XY精蟲

40-80%Percoll分層液中，80%Percoll分層液中X精蟲比率提高 (49->52%)
但40-80%Percoll分層仍無法有效率的分離XY精蟲

http://www.springerlink.com/content/wng5773651076263/

J Assist Reprod Genet. 1998 Oct;15(9):565-9.

Separating X-bearing human spermatozoa through a discontinuous Percoll density gradient proved to be inefficient by double-label fluorescent in situ hybridization.

Lin SP, Lee RK, Tsai YJ, Hwu YM, Lin MH.

Source

Department of Medical Research, Mackay Memorial Hospital, Tamshui, Taipei, Taiwan.

Abstract

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 separatedspermatozoa 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 spermatozoawas 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-bearingspermatozoa is insufficient for clinical use in preconceptional sex selection.