| Development of a cryopreservation protocol for Leydig cells. | |
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MedLine Citation:
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PMID: 17596277 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: In the present study, we describe a procedure to cryopreserve the postnatal members of the Leydig cell lineage, including progenitor (PLC), immature (ILC) and adult (ALC) Leydig cells from, respectively 21-, 35- and 90-day-old rats. METHODS: The cells were resuspended in a culture medium supplemented with 1% bovine serum albumin (Dulbecco's Modified Eagle's Medium [DMEM]/F12) to a final concentration of 2 x 10(6)cells/ml and the effects of varying concentrations of dimethylsulfoxide (DMSO) (5, 10, 15 or 20%) were assessed after freezing at -70 degrees C and then storing in liquid nitrogen. After 12 months of frozen storage, these cells were thawed rapidly at 37 degrees C and Trypan Blue exclusion staining and attachment to culture dishes were assessed as measures of viability. RESULTS: The trypan blue exclusion and attachment rates for Leydig cell stages were around 85% in the presence of 15% DMSO. After frozen storage, Leydig cell steroidogenic capacity in response to a range of LH doses, (0.01-100 ng/ml) was unchanged compared with freshly isolated control cells. Furthermore, the steady-state mRNA levels for Leydig cell specific transcripts were maintained. CONCLUSIONS: This study demonstrates that purified rat Leydig cells at a range of developmental stages can be frozen and that the cryopreserved cells retain normal function. |
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Authors:
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Guo-Rong Chen; Ren-Shan Ge; Han Lin; Lei Dong; Chantal M Sottas; Matthew P Hardy |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2007-06-27 |
Journal Detail:
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Title: Human reproduction (Oxford, England) Volume: 22 ISSN: 0268-1161 ISO Abbreviation: Hum. Reprod. Publication Date: 2007 Aug |
Date Detail:
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Created Date: 2007-07-27 Completed Date: 2007-10-12 Revised Date: 2007-12-03 |
Medline Journal Info:
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Nlm Unique ID: 8701199 Medline TA: Hum Reprod Country: England |
Other Details:
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Languages: eng Pagination: 2160-8 Citation Subset: IM |
Affiliation:
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Department of Pathology of the 1st Affiliated Hospital, Wenzhou Medical College, Wenzhou, Zhejiang 325000, People's Republic of China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Androstane-3,17-diol
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biosynthesis Androsterone / biosynthesis Animals Cell Proliferation Cell Survival Cryopreservation / methods* Dimethyl Sulfoxide / pharmacology Leydig Cells / cytology*, drug effects, metabolism Luteinizing Hormone / pharmacology Male Rats Rats, Sprague-Dawley Testosterone / biosynthesis |
| Grant Support | |
ID/Acronym/Agency:
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HD 33000/HD/NICHD NIH HHS |
| Chemical | |
Reg. No./Substance:
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25126-76-5/Androstane-3,17-diol; 53-41-8/Androsterone; 58-22-0/Testosterone; 67-68-5/Dimethyl Sulfoxide; 9002-67-9/Luteinizing Hormone |
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