Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation
Background: L-proline is a natural, nontoxic cryoprotectant that helps cells and tissues to tolerate freezing in a variety of plants and animals. The use of L-proline in mammalian oocyte cryopreservation is rare. In this study, we explored the cryobiological characteristics of L-proline and evaluate...
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| Format: | Article |
| Language: | English |
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Wolters Kluwer
2016-01-01
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| Series: | Chinese Medical Journal |
| Subjects: | |
| Online Access: | http://www.cmj.org/article.asp?issn=0366-6999;year=2016;volume=129;issue=16;spage=1963;epage=1968;aulast=Zhang |
| _version_ | 1811273239405527040 |
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| author | Lu Zhang Xu Xue Jie Yan Li-Ying Yan Xiao-Hu Jin Xiao-Hui Zhu Zhi-Zhu He Jing Liu Rong Li Jie Qiao |
| author_facet | Lu Zhang Xu Xue Jie Yan Li-Ying Yan Xiao-Hu Jin Xiao-Hui Zhu Zhi-Zhu He Jing Liu Rong Li Jie Qiao |
| author_sort | Lu Zhang |
| collection | DOAJ |
| description | Background: L-proline is a natural, nontoxic cryoprotectant that helps cells and tissues to tolerate freezing in a variety of plants and animals. The use of L-proline in mammalian oocyte cryopreservation is rare. In this study, we explored the cryobiological characteristics of L-proline and evaluated its protective effect in mouse oocyte cryopreservation.
Methods: The freezing property of L-proline was detected by Raman spectroscopy and osmometer. Mature oocytes obtained from 8-week-old B6D2F1 mice were vitrified in a solution consisting various concentration of L-proline with a reduced proportion of dimethyl sulfoxide (DMSO) and ethylene glycol (EG), comparing with the control group (15% DMSO and 15% EG without L-proline). The survival rate, 5-methylcytosine (5-mC) expression, fertilization rate, two-cell rate, and blastocyst rate in vitro were assessed by immunofluorescence and in vitro fertilization. Data were analyzed by Chi-square test.
Results: L-proline can penetrate the oocyte membrane within 1 min. The osmotic pressure of 2.00 mol/L L-proline mixture is similar to that of the control group. The survival rate of the postthawed oocyte in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG is significantly higher than that of the control group. There is no difference of 5-mC expression between the L-proline combination groups and control. The fertilization rate, two-cell rate, and blastocyst rate in vitro from oocyte vitrified in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG solution are similar to that of control.
Conclusions: It indicated that an appropriate concentration of L-proline can improve the cryopreservation efficiency of mouse oocytes with low concentrations of DMSO and EG, which may be applicable to human oocyte vitrification. |
| first_indexed | 2024-04-12T22:54:25Z |
| format | Article |
| id | doaj.art-3c4c92116d954b0788cce5e99af05dfe |
| institution | Directory Open Access Journal |
| issn | 0366-6999 |
| language | English |
| last_indexed | 2024-04-12T22:54:25Z |
| publishDate | 2016-01-01 |
| publisher | Wolters Kluwer |
| record_format | Article |
| series | Chinese Medical Journal |
| spelling | doaj.art-3c4c92116d954b0788cce5e99af05dfe2022-12-22T03:13:13ZengWolters KluwerChinese Medical Journal0366-69992016-01-01129161963196810.4103/0366-6999.187846Cryobiological Characteristics of L-proline in Mammalian Oocyte CryopreservationLu ZhangXu XueJie YanLi-Ying YanXiao-Hu JinXiao-Hui ZhuZhi-Zhu HeJing LiuRong LiJie QiaoBackground: L-proline is a natural, nontoxic cryoprotectant that helps cells and tissues to tolerate freezing in a variety of plants and animals. The use of L-proline in mammalian oocyte cryopreservation is rare. In this study, we explored the cryobiological characteristics of L-proline and evaluated its protective effect in mouse oocyte cryopreservation. Methods: The freezing property of L-proline was detected by Raman spectroscopy and osmometer. Mature oocytes obtained from 8-week-old B6D2F1 mice were vitrified in a solution consisting various concentration of L-proline with a reduced proportion of dimethyl sulfoxide (DMSO) and ethylene glycol (EG), comparing with the control group (15% DMSO and 15% EG without L-proline). The survival rate, 5-methylcytosine (5-mC) expression, fertilization rate, two-cell rate, and blastocyst rate in vitro were assessed by immunofluorescence and in vitro fertilization. Data were analyzed by Chi-square test. Results: L-proline can penetrate the oocyte membrane within 1 min. The osmotic pressure of 2.00 mol/L L-proline mixture is similar to that of the control group. The survival rate of the postthawed oocyte in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG is significantly higher than that of the control group. There is no difference of 5-mC expression between the L-proline combination groups and control. The fertilization rate, two-cell rate, and blastocyst rate in vitro from oocyte vitrified in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG solution are similar to that of control. Conclusions: It indicated that an appropriate concentration of L-proline can improve the cryopreservation efficiency of mouse oocytes with low concentrations of DMSO and EG, which may be applicable to human oocyte vitrification.http://www.cmj.org/article.asp?issn=0366-6999;year=2016;volume=129;issue=16;spage=1963;epage=1968;aulast=ZhangCryoprotective Agentant; Mouse Oocyte; Vitrification |
| spellingShingle | Lu Zhang Xu Xue Jie Yan Li-Ying Yan Xiao-Hu Jin Xiao-Hui Zhu Zhi-Zhu He Jing Liu Rong Li Jie Qiao Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation Chinese Medical Journal Cryoprotective Agentant; Mouse Oocyte; Vitrification |
| title | Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation |
| title_full | Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation |
| title_fullStr | Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation |
| title_full_unstemmed | Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation |
| title_short | Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation |
| title_sort | cryobiological characteristics of l proline in mammalian oocyte cryopreservation |
| topic | Cryoprotective Agentant; Mouse Oocyte; Vitrification |
| url | http://www.cmj.org/article.asp?issn=0366-6999;year=2016;volume=129;issue=16;spage=1963;epage=1968;aulast=Zhang |
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