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Establishment of clival chordoma cell line MUG-CC1 and lymphoblastoid cells as a model for potential new treatment strategies.

Gellner V, Tomazic PV, Lohberger B, Meditz K, Heitzer E, Mokry M, Koele W, Leithner A, Liegl-Atzwanger B, Rinner B - Sci Rep (2016)

Bottom Line: MUG-CC1 is strongly positive for brachyury, cytokeratin, and S100.The cell line showed gains of the entire chromosomes 7, 8, 12, 13, 16, 18, and 20, and high level gains on chromosomes 1q21-1q24 and 17q21-17q25.A new, well-characterized clival chordoma cell line, as well as a non-tumorigenic lymphoblastoid cell line should serve as an in vitro model for the development of potential new treatment strategies for patients suffering from this disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Medical University of Graz, 8036 Graz, Austria.

ABSTRACT
Chordomas are rare malignant tumors that develop from embryonic remnants of the notochord and arise only in the midline from the clivus to the sacrum. Surgery followed by radiotherapy is the standard treatment. As chordomas are resistant to standard chemotherapy, further treatment options are urgently needed. We describe the establishment of a clivus chordoma cell line, MUG-CC1. The cell line is characterized according to its morphology, immunohistochemistry, and growth kinetics. During establishment, cell culture supernatants were collected, and the growth factors HGF, SDF-1, FGF2, and PDGF analyzed using xMAP(®) technology. A spontaneous lymphoblastoid EBV-positive cell line was also developed and characterized. MUG-CC1 is strongly positive for brachyury, cytokeratin, and S100. The cell line showed gains of the entire chromosomes 7, 8, 12, 13, 16, 18, and 20, and high level gains on chromosomes 1q21-1q24 and 17q21-17q25. During cultivation, there was significant expression of HGF and SDF-1 compared to continuous chordoma cell lines. A new, well-characterized clival chordoma cell line, as well as a non-tumorigenic lymphoblastoid cell line should serve as an in vitro model for the development of potential new treatment strategies for patients suffering from this disease.

No MeSH data available.


Related in: MedlinePlus

Growth factors detection by xMAP® technology.Supernatant of chordoma cell lines (MUG-Chor1, U-CH1, U-CH2, U-CH3), human skin fibroblasts (fibro), and tumor surrounding cells (TSC) compared with the clival chordoma MUG-CC1 after 24, 48, and 72 h. Clivus I represents the supernatant after these time points, clivus II the repeated measures after a medium change. (A) Significant expression of HGF in TSC (dashed box) and MUG-CC1 (unbroken box) compared to the other tested cells and (B) SDF-1 in fibros, TSC (dashed box) and MUG-CC1 (unbroken box) were detected.
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f4: Growth factors detection by xMAP® technology.Supernatant of chordoma cell lines (MUG-Chor1, U-CH1, U-CH2, U-CH3), human skin fibroblasts (fibro), and tumor surrounding cells (TSC) compared with the clival chordoma MUG-CC1 after 24, 48, and 72 h. Clivus I represents the supernatant after these time points, clivus II the repeated measures after a medium change. (A) Significant expression of HGF in TSC (dashed box) and MUG-CC1 (unbroken box) compared to the other tested cells and (B) SDF-1 in fibros, TSC (dashed box) and MUG-CC1 (unbroken box) were detected.

Mentions: Within five months, before the cells were separated in two populations, the supernatant was collected to identify important growth factors for establishing a cell culture and to optimize growth media for establishment of further cell lines. Because connective tissue cells were visible during cultivation, we measured HGF, FGF2, SDF-1, and PDGF compared to stable chordoma cell lines (MUG-Chor118 and U-CH1-319) and healthy normal skin fibroblasts (fibro), as well as tumor-surrounding cancer cells (TSC). We detected a time-dependent (24–72 hours) significant increase of HGF in TSC, especially as MUG-CC1 was becoming established, compared to stable chordoma cell lines and fibroblasts (Fig. 4A). There was a significant increase of SDF-1 in primary MUG-CC1, TSC, and in fibroblasts compared to the other cells as well (Fig. 4B). The fluorescent intensity (FI) minus background was presented in all measurements. The FI signals of FGF2 (0–26 FI) and PDGF (0–3.5 FI) were barely detectable or were part of the FI background signal (under the detection limit) and therefore had no effect on the cell cultures (Suppl. Fig. 2).


Establishment of clival chordoma cell line MUG-CC1 and lymphoblastoid cells as a model for potential new treatment strategies.

Gellner V, Tomazic PV, Lohberger B, Meditz K, Heitzer E, Mokry M, Koele W, Leithner A, Liegl-Atzwanger B, Rinner B - Sci Rep (2016)

Growth factors detection by xMAP® technology.Supernatant of chordoma cell lines (MUG-Chor1, U-CH1, U-CH2, U-CH3), human skin fibroblasts (fibro), and tumor surrounding cells (TSC) compared with the clival chordoma MUG-CC1 after 24, 48, and 72 h. Clivus I represents the supernatant after these time points, clivus II the repeated measures after a medium change. (A) Significant expression of HGF in TSC (dashed box) and MUG-CC1 (unbroken box) compared to the other tested cells and (B) SDF-1 in fibros, TSC (dashed box) and MUG-CC1 (unbroken box) were detected.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4829844&req=5

f4: Growth factors detection by xMAP® technology.Supernatant of chordoma cell lines (MUG-Chor1, U-CH1, U-CH2, U-CH3), human skin fibroblasts (fibro), and tumor surrounding cells (TSC) compared with the clival chordoma MUG-CC1 after 24, 48, and 72 h. Clivus I represents the supernatant after these time points, clivus II the repeated measures after a medium change. (A) Significant expression of HGF in TSC (dashed box) and MUG-CC1 (unbroken box) compared to the other tested cells and (B) SDF-1 in fibros, TSC (dashed box) and MUG-CC1 (unbroken box) were detected.
Mentions: Within five months, before the cells were separated in two populations, the supernatant was collected to identify important growth factors for establishing a cell culture and to optimize growth media for establishment of further cell lines. Because connective tissue cells were visible during cultivation, we measured HGF, FGF2, SDF-1, and PDGF compared to stable chordoma cell lines (MUG-Chor118 and U-CH1-319) and healthy normal skin fibroblasts (fibro), as well as tumor-surrounding cancer cells (TSC). We detected a time-dependent (24–72 hours) significant increase of HGF in TSC, especially as MUG-CC1 was becoming established, compared to stable chordoma cell lines and fibroblasts (Fig. 4A). There was a significant increase of SDF-1 in primary MUG-CC1, TSC, and in fibroblasts compared to the other cells as well (Fig. 4B). The fluorescent intensity (FI) minus background was presented in all measurements. The FI signals of FGF2 (0–26 FI) and PDGF (0–3.5 FI) were barely detectable or were part of the FI background signal (under the detection limit) and therefore had no effect on the cell cultures (Suppl. Fig. 2).

Bottom Line: MUG-CC1 is strongly positive for brachyury, cytokeratin, and S100.The cell line showed gains of the entire chromosomes 7, 8, 12, 13, 16, 18, and 20, and high level gains on chromosomes 1q21-1q24 and 17q21-17q25.A new, well-characterized clival chordoma cell line, as well as a non-tumorigenic lymphoblastoid cell line should serve as an in vitro model for the development of potential new treatment strategies for patients suffering from this disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Medical University of Graz, 8036 Graz, Austria.

ABSTRACT
Chordomas are rare malignant tumors that develop from embryonic remnants of the notochord and arise only in the midline from the clivus to the sacrum. Surgery followed by radiotherapy is the standard treatment. As chordomas are resistant to standard chemotherapy, further treatment options are urgently needed. We describe the establishment of a clivus chordoma cell line, MUG-CC1. The cell line is characterized according to its morphology, immunohistochemistry, and growth kinetics. During establishment, cell culture supernatants were collected, and the growth factors HGF, SDF-1, FGF2, and PDGF analyzed using xMAP(®) technology. A spontaneous lymphoblastoid EBV-positive cell line was also developed and characterized. MUG-CC1 is strongly positive for brachyury, cytokeratin, and S100. The cell line showed gains of the entire chromosomes 7, 8, 12, 13, 16, 18, and 20, and high level gains on chromosomes 1q21-1q24 and 17q21-17q25. During cultivation, there was significant expression of HGF and SDF-1 compared to continuous chordoma cell lines. A new, well-characterized clival chordoma cell line, as well as a non-tumorigenic lymphoblastoid cell line should serve as an in vitro model for the development of potential new treatment strategies for patients suffering from this disease.

No MeSH data available.


Related in: MedlinePlus