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Jian-Ping Xu

Publications and source records attributed to Jian-Ping Xu.

21 records · Page 2Linked to original sources

[Isolation and identification of spermatids from mouse testis].

OBJECTIVES: To develop a simple and effective method by which spermatids can be isolated from mouse testis. METHODS: Combination of enzymatic digestion was used to prepare suspension of spermatogenic cells from adult mouse testis, and then a modified discontinuous Percoll gradient (15%, 22%, 30%, 40%, 50%, 60%) centrifugation method was introduced to isolate spermatids from the cellular suspension. The content of spermatids in each isolated fraction by Percoll method was determined by morphology (Wright-Giemsa staining) and flow cytometry analysis, and the viability of spermatogenic cells was assessed using Eosin Y exclusion test. RESULTS: More than 97% of the testicular cells remained their viability after enzymatic digestion. After Percoll centrifuged, six fractions were formed. In each isolated fraction, the 22% fraction contained mostly spermatids(mean 86.7%) and cell viability was more than 85.5%. While in the 30% fraction, immature spermatogenic cells were present, and more than 92% of the cells remained their viability. CONCLUSIONS: A large of relatively purified spermatids can be isolated from mouse testis by enzymatic digestion combined discontinuous Percoll gradient centrifugation method.

Animals↗

[Isolation and identification of spermatids in semen of male infertile patients].

OBJECTIVES: To develop a method by which large and purified populations of spermatids can be isolated in semen of male infertile patients. METHODS: A total of fifteen ejaculates containing cellular elements from infertile patients with various andrological pathologies were obtained after a 24-hour abstinence. A modified discontinuous Percoll gradient (15%, 22%, 30%, 40%, 50%, 60%) centrifugation method was used to isolate the spermatids. After centrifugation at 2,000 r/min for 30 min at 18 degrees C, the single Percoll fractions were separated and analyzed in order to select the one with the greatest purity of spermatid. The germinal cells in each isolated fraction were counted using a Macro sperm counting chamber, then the contents of spermatids were determined by morphology (Wright-Giemsa staining method) and flow cytometry (FCM) analysis, while the contaminated leukocytes were assessed by anti-CD45 immunocytochemistry. RESULTS: After Percoll centrifuged, six single fractions were obtained. Morphology and FCM analysis showed that the 22% fraction contained mostly spermatids [(91.85 +/- 5.18)%, P < 0.005] and the mean density in this fraction was (1.010 +/- 0.786) x 10(5)/ml. While in the 30% fraction, various immature spermatogenic cells including spermatids were present and leukocytes mostly presented in the 60% fraction. CONCLUSIONS: A large population of relatively purified spermatids can be isolated from the ejaculates of infertile patients by using this modified discontinuous Percoll gradient centrifugation method.

Cell Separation↗