Development of a cryopreservation protocol for Leydig cells.

Development of a cryopreservation protocol for Leydig cells. Research Abstract Details 

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Development of a cryopreservation protocol for Leydig cells. Abstract Text:

Guo-Rong Chen,Ren-Shan Ge,Han Lin,Lei Dong,Chantal M Sottas,Matthew P Hardy,

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.

Development of a cryopreservation protocol for Leydig cells. Publishing Authors By Initials

GR Chen,RS Ge,H Lin,L Dong,CM Sottas,MP Hardy,

For similar polycyclic compounds: steroids: androstanes: androstenes: androstenols: testosterone research abstracts see: polycyclic compounds: steroids: androstanes: androstenes: androstenols: testosterone research

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Development of a cryopreservation protocol for Leydig cells. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Human reproduction (Oxford, England)

VOLUME: 22

Page Numbers: 2160-8

Journal Abbreviation: Hum. Reprod.

ISSN: 0268-1161

DAY: 27

MONTH: 06

YEAR: 2007

Development of a cryopreservation protocol for Leydig cells. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 8701199

Development of a cryopreservation protocol for Leydig cells. Keywords Mesh Terms:

KEYWORDS: Testosterone

MESH TERMS: biosynthesis

Chemical & Substance for Abstract: Development of a cryopreservation protocol for Leydig cells. Information

Substance Name: Luteinizing Hormone

Registry Number: 9002-67-9

Grant and Affiliation Information for Development of a cryopreservation protocol for Leydig cells.

AFFILIATION: Department of Pathology of the 1st Affiliated Hospital, Wenzhou Medical College, Wenzhou, Zhejiang 325000, People's Republic of China.

Country: England

AGENCY: United States NICHD

GRANT: HD 33000

ACRONYM: HD

MEDLINETA: Hum Reprod

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