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. 2025 Feb 10;20(2):e0306235.
doi: 10.1371/journal.pone.0306235. eCollection 2025.

The effects of L-carnitine and fructose in improved Ham's F10 on sperm culture in idiopathic severe asthenospermia within 24h

Dehui Chang  1 , Fudong Li  1 , Yindong Kang  1 , Yixin Yan  1 , Feiyan Kong  2 , Wei Jiang  3 , Dongxing Wang  1 , Zhigang Cao  1 , Liuting Xu  1 , Chuang Huang  1 , Yafen Kang  1 , Xuejun Shang  4 , Bin Zhang  1
Affiliations

The effects of L-carnitine and fructose in improved Ham's F10 on sperm culture in idiopathic severe asthenospermia within 24h

Dehui Chang et al. PLoS One. .

Abstract

To study the effects of L-carnitine and fructose on semen parameters of severe asthenospermia patients by sperm culturing in vitro within 24h. We optimized the energy composition and antioxidant substances of sperm culture medium in vitro (based on Ham's F10 culture medium) by orthogonal test for preparing high quality culture medium. Sperms of 60 patients with idiopathic severe asthenospermia were collected, and cultured in vitro within 24h, by Ham's F10 culture medium added to different concentrations of L-carnitine and fructose and culture temperature, whose effects on sperm motility were observed to determine which is the most appropriate concentration and temperature. For determining the appropriate concentration of L-carnitine and fructose and the suitable culture temperature in Ham's F10 culture medium, the orthogonal experiments were carried out to optimize above three factors, which had great influence on sperm viability, survival rate, deformity rate and DNA fragmentation index (DFI). The final concentration of L-carnitine and fructose was determined in terms of initial tests to assess the effects of different concentrations (4, 8, 12, and 16 mg/ml L-carnitine and 0.125, 0.250, 0.375, and 0.50 mg/ml fructose) on sperm viability and motility in culture. During the operation of processing and culturing sperms in vitro within 24h, orthogonal test showed that sperm viability was better at the final concentration of 8 mg/ml L-carnitine and 0.375 mg/ml fructose in improved Ham's F10 culture medium at 36.5°C. Idiopathic severe asthenospermia sperm can be effectively improved by the modified Ham's F10 culture medium of the final concentration of 8 mg/ml L-carnitine and 0.375 mg/ml fructose at 36.5°C within 24h, which has shown better culture effect and is superior to Ham's F10 basic medium.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. All authors also declare that they have agreed to participate in this study and agree to publish the obtained data and analysis results of this study.

Figures

Fig 1
Fig 1. Changes of sperm motility of groups in different concentrations of L-carnitine culture (x¯±s).
Note: *: VS Basic data1, P<0.05.
Fig 2
Fig 2. Changes of sperm motility in different concentrations of fructose culture groups (x¯±s).
Note: *: VS Basic data2, P<0.05.
Fig 3
Fig 3. Effect diagram of orthogonal test.
Fig 4
Fig 4. Changes of semen parameters in groups after 30 min culture.
Note: *: VS Basic data3, P<0.05.
Fig 5
Fig 5. Changes of semen parameters in groups after 1 h culture.
Note: *: VS Basic data3, P<0.05.
Fig 6
Fig 6. Changes of semen parameters in groups after 12 h culture.
Note: *: Basic data3, P<0.05.
Fig 7
Fig 7. Changes of semen parameters between groups after 24h culture.
Note: *: VS Basic data3, P<0.05.
Fig 8
Fig 8. Semen DNA fragmentation index staining of the groups basic data3, NS3, HF3, and m-HF after 30min, 1h, 12h, and 24h culture (light microscope X400).
Note: Semen DNA fragmentation index reflect the degree of sperm DNA integrity and the ultrastructural damage of sperm. The more damaged and fragmented sperms appear, the higher the fragmentation rate increased, which results in poor integrity and less halos on the figures. On the contrary, halo is more with normal sperms. This clinical trial was only to test the efficacy of modified Ham’s F10 medium on semen culture in vitro for patients with idiopathic severe asthenospermia. It has not entered the clinical trial stage for ART assisted pregnancy treatment. Therefore, there was no clinical outcome and harms for the patients involved in the study.
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