Establishing a cryopreservation protocol for patient-derived xenografts of prostate cancer

Laura H. Porter, Mitchell G. Lawrence, Hong Wang, Ashlee K Clark, Andrew Bakshi, Daisuke Obinata, David Goode, Melissa Papargiris, David Clouston, Andrew Ryan, Sam Norden, Eva Corey, Peter S. Nelson, John T. Isaacs, Jeremy Grummet, John Kourambas, Shahneen Sandhu, Declan G. Murphy, David Pook, Mark Frydenberg & 2 others Renea A. Taylor, Gail P. Risbridger

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Background: Serially transplantable patient-derived xenografts (PDXs) are invaluable preclinical models for studying tumor biology and evaluating therapeutic agents. As these models are challenging to establish from prostate cancer specimens, the ability to preserve them through cryopreservation has several advantages for ongoing research. Despite this, there is still uncertainty about the ability to cryopreserve PDXs of prostate cancer. This study compared three different cryopreservation protocols to identify a method that can be used to reproducibly cryopreserve a diverse cohort of prostate cancer PDX models. Methods: One serially transplantable prostate cancer PDX from the Melbourne Urological Research Alliance cohort was used to compare three cryopreservation protocols: slow freezing in fetal calf serum (FCS) with 10% dimethyl sulfoxide (DMSO), FCS with 10% DMSO supplemented with the Rho-associated kinase (ROCK) inhibitor Y-27632 and vitrification. The efficiency of the slow freezing protocols was then assessed in 17 additional prostate cancer PDXs. Following cryopreservation, PDXs were re-established in host mice that were either intact and supplemented with testosterone or castrated. Graft take rate, tumor growth, histological features, and transcriptome profiles before and after cryopreservation were compared. Results: Slow freezing maintained the viability and histological features of prostate cancer PDXs, and the addition of a ROCK inhibitor increased their growth following cryopreservation. Using the slow freezing method, we re-established 100% of PDXs grown in either testosterone-supplemented or castrated host mice. Importantly, the long-term tumor growth rate and transcriptome profile were maintained following cryopreservation. Conclusion: This study has identified a protocol to reliably cryopreserve and re-establish a diverse cohort of serially transplantable PDXs of prostate cancer. This study has the potential to significantly improve the practicality of maintaining PDX models. Cryopreservation may also increase the accessibility of these important resources and provide new opportunities for preclinical studies on a broader spectrum of prostate tumors.

Original languageEnglish
Pages (from-to)1326-1337
Number of pages12
JournalProstate
Volume79
Issue number11
DOIs
Publication statusPublished - 1 Aug 2019

Keywords

  • castration-resistant prostate cancer
  • freezing
  • localized prostate cancer
  • patient-derived xenografts

Cite this

Porter, Laura H. ; Lawrence, Mitchell G. ; Wang, Hong ; Clark, Ashlee K ; Bakshi, Andrew ; Obinata, Daisuke ; Goode, David ; Papargiris, Melissa ; Clouston, David ; Ryan, Andrew ; Norden, Sam ; Corey, Eva ; Nelson, Peter S. ; Isaacs, John T. ; Grummet, Jeremy ; Kourambas, John ; Sandhu, Shahneen ; Murphy, Declan G. ; Pook, David ; Frydenberg, Mark ; Taylor, Renea A. ; Risbridger, Gail P. / Establishing a cryopreservation protocol for patient-derived xenografts of prostate cancer. In: Prostate. 2019 ; Vol. 79, No. 11. pp. 1326-1337.
@article{2560c8b0ab4d4097ad545cbe97c1d718,
title = "Establishing a cryopreservation protocol for patient-derived xenografts of prostate cancer",
abstract = "Background: Serially transplantable patient-derived xenografts (PDXs) are invaluable preclinical models for studying tumor biology and evaluating therapeutic agents. As these models are challenging to establish from prostate cancer specimens, the ability to preserve them through cryopreservation has several advantages for ongoing research. Despite this, there is still uncertainty about the ability to cryopreserve PDXs of prostate cancer. This study compared three different cryopreservation protocols to identify a method that can be used to reproducibly cryopreserve a diverse cohort of prostate cancer PDX models. Methods: One serially transplantable prostate cancer PDX from the Melbourne Urological Research Alliance cohort was used to compare three cryopreservation protocols: slow freezing in fetal calf serum (FCS) with 10{\%} dimethyl sulfoxide (DMSO), FCS with 10{\%} DMSO supplemented with the Rho-associated kinase (ROCK) inhibitor Y-27632 and vitrification. The efficiency of the slow freezing protocols was then assessed in 17 additional prostate cancer PDXs. Following cryopreservation, PDXs were re-established in host mice that were either intact and supplemented with testosterone or castrated. Graft take rate, tumor growth, histological features, and transcriptome profiles before and after cryopreservation were compared. Results: Slow freezing maintained the viability and histological features of prostate cancer PDXs, and the addition of a ROCK inhibitor increased their growth following cryopreservation. Using the slow freezing method, we re-established 100{\%} of PDXs grown in either testosterone-supplemented or castrated host mice. Importantly, the long-term tumor growth rate and transcriptome profile were maintained following cryopreservation. Conclusion: This study has identified a protocol to reliably cryopreserve and re-establish a diverse cohort of serially transplantable PDXs of prostate cancer. This study has the potential to significantly improve the practicality of maintaining PDX models. Cryopreservation may also increase the accessibility of these important resources and provide new opportunities for preclinical studies on a broader spectrum of prostate tumors.",
keywords = "castration-resistant prostate cancer, freezing, localized prostate cancer, patient-derived xenografts",
author = "Porter, {Laura H.} and Lawrence, {Mitchell G.} and Hong Wang and Clark, {Ashlee K} and Andrew Bakshi and Daisuke Obinata and David Goode and Melissa Papargiris and David Clouston and Andrew Ryan and Sam Norden and Eva Corey and Nelson, {Peter S.} and Isaacs, {John T.} and Jeremy Grummet and John Kourambas and Shahneen Sandhu and Murphy, {Declan G.} and David Pook and Mark Frydenberg and Taylor, {Renea A.} and Risbridger, {Gail P.}",
year = "2019",
month = "8",
day = "1",
doi = "10.1002/pros.23839",
language = "English",
volume = "79",
pages = "1326--1337",
journal = "Prostate",
issn = "0270-4137",
publisher = "Wiley-Blackwell",
number = "11",

}

Porter, LH, Lawrence, MG, Wang, H, Clark, AK, Bakshi, A, Obinata, D, Goode, D, Papargiris, M, Clouston, D, Ryan, A, Norden, S, Corey, E, Nelson, PS, Isaacs, JT, Grummet, J, Kourambas, J, Sandhu, S, Murphy, DG, Pook, D, Frydenberg, M, Taylor, RA & Risbridger, GP 2019, 'Establishing a cryopreservation protocol for patient-derived xenografts of prostate cancer', Prostate, vol. 79, no. 11, pp. 1326-1337. https://doi.org/10.1002/pros.23839

Establishing a cryopreservation protocol for patient-derived xenografts of prostate cancer. / Porter, Laura H.; Lawrence, Mitchell G.; Wang, Hong; Clark, Ashlee K; Bakshi, Andrew; Obinata, Daisuke; Goode, David; Papargiris, Melissa; Clouston, David; Ryan, Andrew; Norden, Sam; Corey, Eva; Nelson, Peter S.; Isaacs, John T.; Grummet, Jeremy; Kourambas, John; Sandhu, Shahneen; Murphy, Declan G.; Pook, David; Frydenberg, Mark; Taylor, Renea A.; Risbridger, Gail P.

In: Prostate, Vol. 79, No. 11, 01.08.2019, p. 1326-1337.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Establishing a cryopreservation protocol for patient-derived xenografts of prostate cancer

AU - Porter, Laura H.

AU - Lawrence, Mitchell G.

AU - Wang, Hong

AU - Clark, Ashlee K

AU - Bakshi, Andrew

AU - Obinata, Daisuke

AU - Goode, David

AU - Papargiris, Melissa

AU - Clouston, David

AU - Ryan, Andrew

AU - Norden, Sam

AU - Corey, Eva

AU - Nelson, Peter S.

AU - Isaacs, John T.

AU - Grummet, Jeremy

AU - Kourambas, John

AU - Sandhu, Shahneen

AU - Murphy, Declan G.

AU - Pook, David

AU - Frydenberg, Mark

AU - Taylor, Renea A.

AU - Risbridger, Gail P.

PY - 2019/8/1

Y1 - 2019/8/1

N2 - Background: Serially transplantable patient-derived xenografts (PDXs) are invaluable preclinical models for studying tumor biology and evaluating therapeutic agents. As these models are challenging to establish from prostate cancer specimens, the ability to preserve them through cryopreservation has several advantages for ongoing research. Despite this, there is still uncertainty about the ability to cryopreserve PDXs of prostate cancer. This study compared three different cryopreservation protocols to identify a method that can be used to reproducibly cryopreserve a diverse cohort of prostate cancer PDX models. Methods: One serially transplantable prostate cancer PDX from the Melbourne Urological Research Alliance cohort was used to compare three cryopreservation protocols: slow freezing in fetal calf serum (FCS) with 10% dimethyl sulfoxide (DMSO), FCS with 10% DMSO supplemented with the Rho-associated kinase (ROCK) inhibitor Y-27632 and vitrification. The efficiency of the slow freezing protocols was then assessed in 17 additional prostate cancer PDXs. Following cryopreservation, PDXs were re-established in host mice that were either intact and supplemented with testosterone or castrated. Graft take rate, tumor growth, histological features, and transcriptome profiles before and after cryopreservation were compared. Results: Slow freezing maintained the viability and histological features of prostate cancer PDXs, and the addition of a ROCK inhibitor increased their growth following cryopreservation. Using the slow freezing method, we re-established 100% of PDXs grown in either testosterone-supplemented or castrated host mice. Importantly, the long-term tumor growth rate and transcriptome profile were maintained following cryopreservation. Conclusion: This study has identified a protocol to reliably cryopreserve and re-establish a diverse cohort of serially transplantable PDXs of prostate cancer. This study has the potential to significantly improve the practicality of maintaining PDX models. Cryopreservation may also increase the accessibility of these important resources and provide new opportunities for preclinical studies on a broader spectrum of prostate tumors.

AB - Background: Serially transplantable patient-derived xenografts (PDXs) are invaluable preclinical models for studying tumor biology and evaluating therapeutic agents. As these models are challenging to establish from prostate cancer specimens, the ability to preserve them through cryopreservation has several advantages for ongoing research. Despite this, there is still uncertainty about the ability to cryopreserve PDXs of prostate cancer. This study compared three different cryopreservation protocols to identify a method that can be used to reproducibly cryopreserve a diverse cohort of prostate cancer PDX models. Methods: One serially transplantable prostate cancer PDX from the Melbourne Urological Research Alliance cohort was used to compare three cryopreservation protocols: slow freezing in fetal calf serum (FCS) with 10% dimethyl sulfoxide (DMSO), FCS with 10% DMSO supplemented with the Rho-associated kinase (ROCK) inhibitor Y-27632 and vitrification. The efficiency of the slow freezing protocols was then assessed in 17 additional prostate cancer PDXs. Following cryopreservation, PDXs were re-established in host mice that were either intact and supplemented with testosterone or castrated. Graft take rate, tumor growth, histological features, and transcriptome profiles before and after cryopreservation were compared. Results: Slow freezing maintained the viability and histological features of prostate cancer PDXs, and the addition of a ROCK inhibitor increased their growth following cryopreservation. Using the slow freezing method, we re-established 100% of PDXs grown in either testosterone-supplemented or castrated host mice. Importantly, the long-term tumor growth rate and transcriptome profile were maintained following cryopreservation. Conclusion: This study has identified a protocol to reliably cryopreserve and re-establish a diverse cohort of serially transplantable PDXs of prostate cancer. This study has the potential to significantly improve the practicality of maintaining PDX models. Cryopreservation may also increase the accessibility of these important resources and provide new opportunities for preclinical studies on a broader spectrum of prostate tumors.

KW - castration-resistant prostate cancer

KW - freezing

KW - localized prostate cancer

KW - patient-derived xenografts

UR - http://www.scopus.com/inward/record.url?scp=85068092094&partnerID=8YFLogxK

U2 - 10.1002/pros.23839

DO - 10.1002/pros.23839

M3 - Article

VL - 79

SP - 1326

EP - 1337

JO - Prostate

JF - Prostate

SN - 0270-4137

IS - 11

ER -