Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles

Lea Bejstrup Jensen; Cristina Subiran Adrados; Jane Alrø Bøtkjær; Jesús Cadenas; Sivanandane Sittadjody; Emmanuel Opara; Pernille Landbæk Sørensen; Kirsten Tryde Macklon; Anette Tønnes Pedersen; Stine Gry Kristensen

doi:10.1210/clinem/dgaf218

Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles

Lea Bejstrup Jensen^*, Cristina Subiran Adrados, Jane Alrø Bøtkjær, Jesús Cadenas, Sivanandane Sittadjody, Emmanuel Opara, Pernille Landbæk Sørensen, Kirsten Tryde Macklon, Anette Tønnes Pedersen, Stine Gry Kristensen

^*Corresponding author for this work

Department of Gynaecology, Fertility and Births

3 Citations (Scopus)

Abstract

CONTEXT: The transformation of follicular granulosa cells into luteal cells of the corpus luteum remains poorly understood in the human ovary.

OBJECTIVE: To investigate the luteinization process and steroidogenic differences between granulosa cells from small antral and preovulatory follicles in vitro.

METHODS: At the University Hospital of Copenhagen, Denmark, and Wake Forest Institute for Regenerative Medicine, USA, granulosa-lutein cells were obtained from 12 women undergoing IVF treatment, while follicular granulosa cells from unstimulated small antral follicles and corpus luteum were collected from 18 women undergoing ovarian tissue cryopreservation. Cells were cultured for up to 96 hours or 12 days with or without androstenedione or testosterone supplementation and analyzed using RT-qPCR and steroid hormone assays.

RESULTS: In follicular granulosa cells, luteinization markers (CYP11A1, P < .05; STAR, P < .001) increased within 24 to 48 hours, while granulosa markers (HSD17β1, P < .001; CYP19A1, P < .05) decreased within 6 to 12 hours. Luteinizing hormone/choriogonadotropin receptor remained unchanged. By 48 hours, gene expression resembled that of the corpus luteum. In contrast, granulosa-lutein cells exhibited highly luteinized profiles from day 0, with significantly higher progesterone/(17)estradiol ratios. Androgen supplementation and long-term follicle-stimulating hormone exposure did not alter luteinization.

CONCLUSION: This study uniquely demonstrates that unstimulated follicular granulosa cells undergo a gradual, intrinsic luteinization process, independent of external hormonal triggers. In contrast, granulosa-lutein cells are already highly luteinized upon aspiration. These findings challenge conventional views on luteinization and highlight intrinsic cellular programming as a key driver, offering new insights into ovarian physiology and potential therapeutic targets for reproductive disorders.

Original language	English
Journal	The Journal of clinical endocrinology and metabolism
Volume	110
Issue number	12
Pages (from-to)	3470-3481
Number of pages	12
ISSN	0021-972X
DOIs	https://doi.org/10.1210/clinem/dgaf218
Publication status	Published - 18 Nov 2025

Keywords

granulosa cells
hormone secretion
luteinization
preovulatory follicles
small antral follicles
timeline

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10.1210/clinem/dgaf218Licence: CC BY

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Jensen, L. B., Adrados, C. S., Bøtkjær, J. A., Cadenas, J., Sittadjody, S., Opara, E., Sørensen, P. L., Macklon, K. T., Pedersen, A. T., & Kristensen, S. G. (2025). Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles. The Journal of clinical endocrinology and metabolism, 110(12), 3470-3481. https://doi.org/10.1210/clinem/dgaf218

Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles. / Jensen, Lea Bejstrup ; Adrados, Cristina Subiran ; Bøtkjær, Jane Alrø et al.
In: The Journal of clinical endocrinology and metabolism, Vol. 110, No. 12, 18.11.2025, p. 3470-3481.

Research output: Contribution to journal › Journal article › Research › peer-review

Jensen, LB , Adrados, CS , Bøtkjær, JA , Cadenas, J, Sittadjody, S, Opara, E, Sørensen, PL , Macklon, KT , Pedersen, AT & Kristensen, SG 2025, 'Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles', The Journal of clinical endocrinology and metabolism, vol. 110, no. 12, pp. 3470-3481. https://doi.org/10.1210/clinem/dgaf218

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title = "Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles",

abstract = "CONTEXT: The transformation of follicular granulosa cells into luteal cells of the corpus luteum remains poorly understood in the human ovary.OBJECTIVE: To investigate the luteinization process and steroidogenic differences between granulosa cells from small antral and preovulatory follicles in vitro.METHODS: At the University Hospital of Copenhagen, Denmark, and Wake Forest Institute for Regenerative Medicine, USA, granulosa-lutein cells were obtained from 12 women undergoing IVF treatment, while follicular granulosa cells from unstimulated small antral follicles and corpus luteum were collected from 18 women undergoing ovarian tissue cryopreservation. Cells were cultured for up to 96 hours or 12 days with or without androstenedione or testosterone supplementation and analyzed using RT-qPCR and steroid hormone assays.RESULTS: In follicular granulosa cells, luteinization markers (CYP11A1, P < .05; STAR, P < .001) increased within 24 to 48 hours, while granulosa markers (HSD17β1, P < .001; CYP19A1, P < .05) decreased within 6 to 12 hours. Luteinizing hormone/choriogonadotropin receptor remained unchanged. By 48 hours, gene expression resembled that of the corpus luteum. In contrast, granulosa-lutein cells exhibited highly luteinized profiles from day 0, with significantly higher progesterone/(17)estradiol ratios. Androgen supplementation and long-term follicle-stimulating hormone exposure did not alter luteinization.CONCLUSION: This study uniquely demonstrates that unstimulated follicular granulosa cells undergo a gradual, intrinsic luteinization process, independent of external hormonal triggers. In contrast, granulosa-lutein cells are already highly luteinized upon aspiration. These findings challenge conventional views on luteinization and highlight intrinsic cellular programming as a key driver, offering new insights into ovarian physiology and potential therapeutic targets for reproductive disorders.",

keywords = "granulosa cells, hormone secretion, luteinization, preovulatory follicles, small antral follicles, timeline",

author = "Jensen, \{Lea Bejstrup\} and Adrados, \{Cristina Subiran\} and B{\o}tkj{\ae}r, \{Jane Alr{\o}\} and Jes{\'u}s Cadenas and Sivanandane Sittadjody and Emmanuel Opara and S{\o}rensen, \{Pernille Landb{\ae}k\} and Macklon, \{Kirsten Tryde\} and Pedersen, \{Anette T{\o}nnes\} and Kristensen, \{Stine Gry\}",

note = "{\textcopyright} The Author(s) 2025. Published by Oxford University Press on behalf of the Endocrine Society.",

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day = "18",

doi = "10.1210/clinem/dgaf218",

language = "English",

volume = "110",

pages = "3470--3481",

journal = "The Journal of clinical endocrinology and metabolism",

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T1 - Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles

AU - Jensen, Lea Bejstrup

AU - Adrados, Cristina Subiran

AU - Bøtkjær, Jane Alrø

AU - Cadenas, Jesús

AU - Sittadjody, Sivanandane

AU - Opara, Emmanuel

AU - Sørensen, Pernille Landbæk

AU - Macklon, Kirsten Tryde

AU - Pedersen, Anette Tønnes

AU - Kristensen, Stine Gry

PY - 2025/11/18

Y1 - 2025/11/18

N2 - CONTEXT: The transformation of follicular granulosa cells into luteal cells of the corpus luteum remains poorly understood in the human ovary.OBJECTIVE: To investigate the luteinization process and steroidogenic differences between granulosa cells from small antral and preovulatory follicles in vitro.METHODS: At the University Hospital of Copenhagen, Denmark, and Wake Forest Institute for Regenerative Medicine, USA, granulosa-lutein cells were obtained from 12 women undergoing IVF treatment, while follicular granulosa cells from unstimulated small antral follicles and corpus luteum were collected from 18 women undergoing ovarian tissue cryopreservation. Cells were cultured for up to 96 hours or 12 days with or without androstenedione or testosterone supplementation and analyzed using RT-qPCR and steroid hormone assays.RESULTS: In follicular granulosa cells, luteinization markers (CYP11A1, P < .05; STAR, P < .001) increased within 24 to 48 hours, while granulosa markers (HSD17β1, P < .001; CYP19A1, P < .05) decreased within 6 to 12 hours. Luteinizing hormone/choriogonadotropin receptor remained unchanged. By 48 hours, gene expression resembled that of the corpus luteum. In contrast, granulosa-lutein cells exhibited highly luteinized profiles from day 0, with significantly higher progesterone/(17)estradiol ratios. Androgen supplementation and long-term follicle-stimulating hormone exposure did not alter luteinization.CONCLUSION: This study uniquely demonstrates that unstimulated follicular granulosa cells undergo a gradual, intrinsic luteinization process, independent of external hormonal triggers. In contrast, granulosa-lutein cells are already highly luteinized upon aspiration. These findings challenge conventional views on luteinization and highlight intrinsic cellular programming as a key driver, offering new insights into ovarian physiology and potential therapeutic targets for reproductive disorders.

AB - CONTEXT: The transformation of follicular granulosa cells into luteal cells of the corpus luteum remains poorly understood in the human ovary.OBJECTIVE: To investigate the luteinization process and steroidogenic differences between granulosa cells from small antral and preovulatory follicles in vitro.METHODS: At the University Hospital of Copenhagen, Denmark, and Wake Forest Institute for Regenerative Medicine, USA, granulosa-lutein cells were obtained from 12 women undergoing IVF treatment, while follicular granulosa cells from unstimulated small antral follicles and corpus luteum were collected from 18 women undergoing ovarian tissue cryopreservation. Cells were cultured for up to 96 hours or 12 days with or without androstenedione or testosterone supplementation and analyzed using RT-qPCR and steroid hormone assays.RESULTS: In follicular granulosa cells, luteinization markers (CYP11A1, P < .05; STAR, P < .001) increased within 24 to 48 hours, while granulosa markers (HSD17β1, P < .001; CYP19A1, P < .05) decreased within 6 to 12 hours. Luteinizing hormone/choriogonadotropin receptor remained unchanged. By 48 hours, gene expression resembled that of the corpus luteum. In contrast, granulosa-lutein cells exhibited highly luteinized profiles from day 0, with significantly higher progesterone/(17)estradiol ratios. Androgen supplementation and long-term follicle-stimulating hormone exposure did not alter luteinization.CONCLUSION: This study uniquely demonstrates that unstimulated follicular granulosa cells undergo a gradual, intrinsic luteinization process, independent of external hormonal triggers. In contrast, granulosa-lutein cells are already highly luteinized upon aspiration. These findings challenge conventional views on luteinization and highlight intrinsic cellular programming as a key driver, offering new insights into ovarian physiology and potential therapeutic targets for reproductive disorders.

KW - granulosa cells

KW - hormone secretion

KW - luteinization

KW - preovulatory follicles

KW - small antral follicles

KW - timeline

UR - https://www.scopus.com/pages/publications/105021950908

U2 - 10.1210/clinem/dgaf218

DO - 10.1210/clinem/dgaf218

M3 - Journal article

C2 - 40186482

SN - 0021-972X

VL - 110

SP - 3470

EP - 3481

JO - The Journal of clinical endocrinology and metabolism

JF - The Journal of clinical endocrinology and metabolism

IS - 12

ER -

Distinct Luteinization Profiles of Cultured Human Granulosa Cells from Small Antral and Preovulatory Follicles

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