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Biobanking of patient and patient-derived xenograft ovarian tumour tissue: efficient preservation with low and high fetal calf serum based methods.

Alkema NG, Tomar T, Duiker EW, Jan Meersma G, Klip H, van der Zee AG, Wisman GB, de Jong S - Sci Rep (2015)

Bottom Line: We successfully established 45 subcutaneous ovarian cancer PDXs, reflecting all histological subtypes, with an overall take rate of 68%.Our results indicate that both protocols can be used for biobanking of ovarian tumour and PDX tissues.Moreover, primary engraftment of fresh patient-derived tumours in mice followed by freezing tissue of successfully established PDXs is the preferred way of efficient ovarian cancer PDX biobanking.

View Article: PubMed Central - PubMed

Affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.

ABSTRACT
Using patient-derived xenografts (PDXs) for preclinical cancer research demands proper storage of tumour material to facilitate logistics and to reduce the number of animals needed. We successfully established 45 subcutaneous ovarian cancer PDXs, reflecting all histological subtypes, with an overall take rate of 68%. Corresponding cells from mouse replaced human tumour stromal and endothelial cells in second generation PDXs as demonstrated with mouse-specific vimentin and CD31 immunohistochemical staining. For biobanking purposes two cryopreservation methods, a fetal calf serum (FCS)-based (95%v/v) "FCS/DMSO" protocol and a low serum-based (10%v/v) "vitrification" protocol were tested. After primary cryopreservation, tumour take rates were 38% and 67% using either the vitrification or FCS/DMSO-based cryopreservation protocol, respectively. Cryopreserved tumour tissue of established PDXs achieved take rates of 67% and 94%, respectively compared to 91% using fresh PDX tumour tissue. Genotyping analysis showed that no changes in copy number alterations were introduced by any of the biobanking methods. Our results indicate that both protocols can be used for biobanking of ovarian tumour and PDX tissues. However, FCS/DMSO-based cryopreservation is more successful. Moreover, primary engraftment of fresh patient-derived tumours in mice followed by freezing tissue of successfully established PDXs is the preferred way of efficient ovarian cancer PDX biobanking.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemistry of the ER and PR in representative slides of the primary tumour tissue of patient 56 (Patient), tumour tissue from F2 generation of PDX 56 (F2), and tumour tissue from F2 generation of cryopreserved PDX 56 F1 using either FCS/DMSO (F2:FCS) or vitrification (F2:Vitri).Magnification 10× and 40×.
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f3: Immunohistochemistry of the ER and PR in representative slides of the primary tumour tissue of patient 56 (Patient), tumour tissue from F2 generation of PDX 56 (F2), and tumour tissue from F2 generation of cryopreserved PDX 56 F1 using either FCS/DMSO (F2:FCS) or vitrification (F2:Vitri).Magnification 10× and 40×.

Mentions: The oestrogen receptor (ER) and progesterone receptor (PR) are known to be frequently expressed in serous ovarian cancer and are also associated with improved survival21. Because of the use of a high percentage of FCS, we hypothesized that FCS containing growth factors may induce a selection on hormone dependent cancer cells and/or changes in signalling pathways. Therefore, we compared the expression of ER and PR between the two storage methods in tumour material from three high-grade serous patients (37, 56 and 67) and one mixed histology ovarian cancer patient (61). Figure 3 shows the expression of ER and PR in tumour tissue from patient 56, tumour tissue from F2 generation of PDX 56, and tumour tissue from F2 generation of cryopreserved PDX 56 generation F1, using either FCS/DMSO or vitrification (Fig. 3). PR expression increased in F2 generations compared with the primary tumour. Neither of the storage methods influenced the expression levels of ER and PR (Fig. 3). In all 4 PDX models, both consistently positive (37, 56 and 67) or negative (61) ER and PR staining was observed through generations.


Biobanking of patient and patient-derived xenograft ovarian tumour tissue: efficient preservation with low and high fetal calf serum based methods.

Alkema NG, Tomar T, Duiker EW, Jan Meersma G, Klip H, van der Zee AG, Wisman GB, de Jong S - Sci Rep (2015)

Immunohistochemistry of the ER and PR in representative slides of the primary tumour tissue of patient 56 (Patient), tumour tissue from F2 generation of PDX 56 (F2), and tumour tissue from F2 generation of cryopreserved PDX 56 F1 using either FCS/DMSO (F2:FCS) or vitrification (F2:Vitri).Magnification 10× and 40×.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4594124&req=5

f3: Immunohistochemistry of the ER and PR in representative slides of the primary tumour tissue of patient 56 (Patient), tumour tissue from F2 generation of PDX 56 (F2), and tumour tissue from F2 generation of cryopreserved PDX 56 F1 using either FCS/DMSO (F2:FCS) or vitrification (F2:Vitri).Magnification 10× and 40×.
Mentions: The oestrogen receptor (ER) and progesterone receptor (PR) are known to be frequently expressed in serous ovarian cancer and are also associated with improved survival21. Because of the use of a high percentage of FCS, we hypothesized that FCS containing growth factors may induce a selection on hormone dependent cancer cells and/or changes in signalling pathways. Therefore, we compared the expression of ER and PR between the two storage methods in tumour material from three high-grade serous patients (37, 56 and 67) and one mixed histology ovarian cancer patient (61). Figure 3 shows the expression of ER and PR in tumour tissue from patient 56, tumour tissue from F2 generation of PDX 56, and tumour tissue from F2 generation of cryopreserved PDX 56 generation F1, using either FCS/DMSO or vitrification (Fig. 3). PR expression increased in F2 generations compared with the primary tumour. Neither of the storage methods influenced the expression levels of ER and PR (Fig. 3). In all 4 PDX models, both consistently positive (37, 56 and 67) or negative (61) ER and PR staining was observed through generations.

Bottom Line: We successfully established 45 subcutaneous ovarian cancer PDXs, reflecting all histological subtypes, with an overall take rate of 68%.Our results indicate that both protocols can be used for biobanking of ovarian tumour and PDX tissues.Moreover, primary engraftment of fresh patient-derived tumours in mice followed by freezing tissue of successfully established PDXs is the preferred way of efficient ovarian cancer PDX biobanking.

View Article: PubMed Central - PubMed

Affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.

ABSTRACT
Using patient-derived xenografts (PDXs) for preclinical cancer research demands proper storage of tumour material to facilitate logistics and to reduce the number of animals needed. We successfully established 45 subcutaneous ovarian cancer PDXs, reflecting all histological subtypes, with an overall take rate of 68%. Corresponding cells from mouse replaced human tumour stromal and endothelial cells in second generation PDXs as demonstrated with mouse-specific vimentin and CD31 immunohistochemical staining. For biobanking purposes two cryopreservation methods, a fetal calf serum (FCS)-based (95%v/v) "FCS/DMSO" protocol and a low serum-based (10%v/v) "vitrification" protocol were tested. After primary cryopreservation, tumour take rates were 38% and 67% using either the vitrification or FCS/DMSO-based cryopreservation protocol, respectively. Cryopreserved tumour tissue of established PDXs achieved take rates of 67% and 94%, respectively compared to 91% using fresh PDX tumour tissue. Genotyping analysis showed that no changes in copy number alterations were introduced by any of the biobanking methods. Our results indicate that both protocols can be used for biobanking of ovarian tumour and PDX tissues. However, FCS/DMSO-based cryopreservation is more successful. Moreover, primary engraftment of fresh patient-derived tumours in mice followed by freezing tissue of successfully established PDXs is the preferred way of efficient ovarian cancer PDX biobanking.

No MeSH data available.


Related in: MedlinePlus