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Culture of human ov...
Culture of human ovarian tissue in xeno-free conditions using laminin components of the human ovarian extracellular matrix
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- Hao, J. (författare)
- Central South University, China; Karolinska Institute, Sweden; Karolinska University Hospital, Sweden; Clinical Research Center for Women’s Reproductive Health in Hunan province, China
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- Tuck, A. R. (författare)
- Karolinska Institute, Sweden; Karolinska University Hospital, Sweden
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- Prakash, C. R. (författare)
- Karolinska Institutet,Karolinska Institute, Sweden; Karolinska University Hospital, Sweden
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- Damdimopoulos, A. (författare)
- Karolinska Institutet,Karolinska Institute, Sweden
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- Sjödin, Marcus (författare)
- RISE,Kemiska processer och läkemedel,Karolinska Institute, Sweden
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- Lindberg, Johan (författare)
- RISE,Kemiska processer och läkemedel,Karolinska Institute, Sweden
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- Niklasson, Boel (författare)
- Sophiahemmet Högskola,Karolinska Institute, Sweden; Sophiahemmet University, Sweden
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- Pettersson, K. (författare)
- Karolinska Institutet,Karolinska Institute, Sweden; Karolinska University Hospital, Sweden
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- Hovatta, O. (författare)
- Karolinska Institute, Sweden; Karolinska University Hospital, Sweden
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- Damdimopoulou, P. (författare)
- Karolinska Institutet,Karolinska Institute, Sweden; Karolinska University Hospital, Sweden
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(creator_code:org_t)
- 2020-07-15
- 2020
- Engelska.
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Ingår i: Journal of Assisted Reproduction and Genetics. - : Springer. - 1058-0468 .- 1573-7330. ; 37, s. 2137-2150
- Relaterad länk:
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https://www.ncbi.nlm...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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http://kipublication...
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Abstract
Ämnesord
Stäng
- Purpose: Our purpose was to identify human ovarian extracellular matrix (ECM) components that would support in vitro culture of human ovarian tissue and be compatible with possible future clinical applications. We characterized ovarian expression of laminins and selected three laminin tripeptides for culture experiments to be compared with Matrigel, an undefined and animal-based mixture of ECM components. Methods: Expression of the 12 laminin genes was determined on transcript and protein levels using cortical tissue samples (n = 6), commercial ovary RNA (n = 1), follicular fluid granulosa cells (n = 20), and single-cell RNA-sequencing data. Laminin 221 (LN221), LN521, LN511, and their mixture were chosen for a 7-day culture experiment along with Matrigel using tissue from 17 patients. At the end of the culture, follicles were evaluated by scoring and counting from serial tissue sections, apoptosis measured using in situ TUNEL assay, proliferation by Ki67 staining, and endocrine function by quantifying steroids in culture media using UPLC-MS/MS. Results: Approximately half of the cells in ovarian cortex expressed at least one laminin gene. The overall most expressed laminin α-chains were LAMA2 and LAMA5, β-chains LAMB1 and LAMB2, and γ-chain LAMC1. In culture experiments, LN221 enhanced follicular survival compared with Matrigel (p < 0.001), whereas tissue cultured on LN521 had higher proportion of secondary follicles (p < 0.001). LN511 and mixture of laminins did not support the cultures leading to lower follicle densities and higher apoptosis. All cultures produced steroids and contained proliferating cells. Conclusions: LN221 and LN521 show promise in providing xeno-free growth substrates for human ovarian tissue cultures, which may help in further development of folliculogenesis in vitro for clinical practices. The system could also be used for identification of adverse effects of chemicals in ovaries.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Reproduktionsmedicin och gynekologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Obstetrics, Gynaecology and Reproductive Medicine (hsv//eng)
Nyckelord
- Human ovary
- Laminins
- Matrigel
- Tissue culture
- Xeno-free
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Hao, J.
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Tuck, A. R.
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Prakash, C. R.
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Damdimopoulos, A ...
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Sjödin, Marcus
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Lindberg, Johan
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visa fler...
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Niklasson, Boel
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Pettersson, K.
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Hovatta, O.
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Damdimopoulou, P ...
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RISE
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Sophiahemmet Högskola
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Karolinska Institutet