人精液中未成熟生殖细胞的检测及临床意义

Determination and Clinical Significance of Immature Germ Cells in Human Semen

目的 :探讨检测人精液中未成熟生殖细胞含量的临床意义。 方法 :运用非连续梯度Percoll法分离人精液中的各类细胞 ,May Grunwald Giemsa染色及FITC标记的CD4 5单克隆抗体免疫荧光鉴定未成熟生殖细胞及白细胞 ,并检测 30份精液中未成熟生殖细胞含量 ,其中生育组 10例 ,不育组 2 0例。 结果 :精液中未成熟生殖细胞主要集中分布于 30 %～ 4 5 %Percoll分离液中 ,白细胞则分布在 5 0 %～ 5 5 %的分离液中。未成熟生殖细胞在生育组为(0 .70± 0 .4 0 )× 10 6/ml,不育组为 (1.2 8± 0 .70 )× 10 6/ml,两组差异有显著性 (P <0 .0 5 ) ,与精子密度、活率及正常形态百分率无明显相关性 (密度r=0 .0 0 2 ,P >0 .0 5 ;活率r=- 0 .2 2 7,P >0 .0 5 ;正常形态百分率r=0 .2 4 0 ,P >0 .0 5 )。 结论 :运用非连续梯度Percoll法能有效地分离精液中未成熟的生殖细胞 ,其含量有可能作为一项临床指标反映精液质量。

Objective: To discuss the clinical significance of determining immature germ cells (IGC) in human semen. Methods: Discontinuous Percoll gradients technique was employed to separate different cells and May-Grunwald-Giemsa staining and fluorescein isothiocyanate (FITC)-Mab-CD45 was adopted to identify IGCs and leukcocytes in semen. The IGCs in 30 semen samples were determined including 10 fertile and 20 infertile cases. Results: IGCs concentrated in gradient fractions with 30% to 45% Percoll and leukocytes concentrated in 50%～55% Percoll fractions. The concentration of IGCs was ( 0.70 ± 0.40 )×10 6 ml in the fertile group and ( 1.28 ± 0.70 )×10 6 /ml in the infertile group(P< 0.05 ). There was no statistical correlation between the IGC concentration and the sperm density, vitality and normal morphology(P> 0.05 ). Conclusion: The use of the discontinuous Percoll gradient method can reach the best separation of IGCs in the ejaculate and it is possible to be used as a clinical index to reflect semen quality.