Single layer centrifugation combined with brief abstinence for semen cryopreservation in a patient with hematospermia: Case report and literature review

Abstract

Rationale: Efficient semen preservation is important in patients with hematospermia (HS). Few reports have delineated effective methods for the removal of red blood cells (RBCs), and existing techniques have exhibited constraints that affect sperm preservation. There is a paucity of studies on semen preservation in HS patients.

Patient concerns: Prior to surgical intervention for an ejaculatory duct cyst, the patient expressed concerns regarding potential fertility impairment. Considering his intention to conceive in the near future, he opted for sperm cryopreservation in a sperm bank as a precautionary measure.

Diagnoses: We report the case of a patient who presented with HS and was diagnosed with an ejaculatory duct cyst accompanied by cyst wall calcification. Semen analysis revealed the presence of numerous RBCs in the ejaculate.

Interventions: Three semen processing techniques to eliminate blood contamination were evaluated in the laboratory. (A) Processing of the RBC lysis buffer, (B) density gradient centrifugation, and (C) single layer centrifugation (SLC). Donor semen mixed with peripheral blood was used for the experiment. Post-thawing progressive motility (%) of the sperm was evaluated using computer-assisted sperm analysis. Additionally, the patient was advised to undergo a brief period of abstinence (4 hours) to diminish the quantity of RBCs in the semen and to facilitate the freezing of additional straws.

Outcomes: The SLC can effectively remove RBCs from semen while maintaining high sperm recovery and post-thawing progressive motility. The combination of SLC and very short-term abstinence elevates the quantity of frozen sperm relative to alternative compositions. Ultimately, the patient successfully cryopreserved 5 semen straws.

Lessons: Before commencing formal treatment, it is essential to evaluate fertility preservation in individuals with HS. Employing SLC and subsequent semen collection following a brief abstinence period can mitigate the adverse effects of physical damage on sperm and avert the influence of RBC lysis on the efficacy of sperm cryopreservation. This method offers an effective strategy for autologous semen cryopreservation in a clinical setting for patients with HS.

Keywords: ejaculatory duct cyst; hematospermia; single layer centrifugation; sperm cryopreservation.

Figure 1.

Schematic illustration for the removal of red blood cells (RBCs) in semen. The RBCs in the samples were removed by 3 different methods: RBC lysis buffer treatment, density gradient centrifugation (DGC), and single layer centrifugation (SLC). After freezing and thawing, sperm concentration and motility parameters were measured by computer-assisted sperm analysis (CASA). The figure was created with BioRender.

Figure 2.

The computer-assisted sperm analysis (CASA) results for the patient with hematospermia. (A and B) The semen from the patient with hematospermia appears bright red, and a high concentration of red blood cells is observed under microscopy. (C and D) Pre- and post-thawing sperm motility parameters were determined using CASA. Sperm motility levels are shown with different colors: red (grade A), blue (grade B), purple (immobile sperm), and yellow (grade D). The scale bar represents 50 μm.

Figure 3.

Comparison of motile sperm recovery across 3 experimental treatments for hematospermia. (A) Red blood cells were incubated with semen to simulate hematospermia formation. Computer-assisted sperm analysis (CASA) results for (B) red blood cell (RBC) lysis treatment, (C) density gradient centrifugation (DGC), and (D) single layer centrifugation. Sperm motility levels are shown with different colors: red (grade A), blue (grade B), purple (grade C), and yellow (immobile sperm). The scale bar represents 50 μm.

Figure 4.

Assessment of red blood cell removal and sperm cryopreservation efficacy in hematospermia. (A) In the semen sample obtained during the first ejaculation, microscopic examination revealed a large number of red blood cells (RBCs) and their fragments. (B and C) Sperm parameters after single layer centrifugation and post-thawing motility were determined using computer-assisted sperm analysis (CASA). (D) After 4 hours of abstinence, the sperm and RBC concentrations in the semen of patients decreased. (E and F) CASA results for the second freezing sperm parameters for SLC and post-thawing motility. Sperm motility levels are shown with different colors: red (grade A), blue (grade B), purple (grade C), and yellow (immobile sperm). The scale bar represents 50 μm.