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Spontaneous generation of germline characteristics in mouse fibrosarcoma cells.

Ma Z, Hu Y, Jiang G, Hou J, Liu R, Lu Y, Liu C - Sci Rep (2012)

Bottom Line: In this study, we further showed that cells functioning as germline could be present in mouse fibrosarcoma cells (L929 cell line).Early germline-like cells spontaneously appeared in L929 cells and further differentiated into oocyte-like cells.These germline-like cells can, in turn, develop into blastocyst-like structures in vitro and cause teratocarcinomas in vivo, which is consistent with natural germ cells in function.

View Article: PubMed Central - PubMed

Affiliation: Department of Labratory Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai 200040, China.

ABSTRACT
Germline/embryonic-specific genes have been found to be activated in somatic tumors. In this study, we further showed that cells functioning as germline could be present in mouse fibrosarcoma cells (L929 cell line). Early germline-like cells spontaneously appeared in L929 cells and further differentiated into oocyte-like cells. These germline-like cells can, in turn, develop into blastocyst-like structures in vitro and cause teratocarcinomas in vivo, which is consistent with natural germ cells in function. Generation of germline-like cells from somatic tumors might provide a novel way to understand why somatic cancer cells have strong features of embryonic/germline development. It is thought that the germline traits of tumors are associated with the central characteristics of malignancy, such as immortalization, invasion, migration and immune evasion. Therefore, germline-like cells in tumors might provide potential targets to tumor biology, diagnosis and therapy.

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Generation of germline-like cells from a single cell.(a) Morphology of a single L929 cell in culture. (b) The isolated L929 cells showed fibroblast-shaped cells at about day 2. (c) Morphology of the single clone at day 5. (d) Round cells appeared above fibroblast-shaped cells in single-cell culture at day 8. (e-h) The cultures were observed at day 16. (e) Round-shaped cells migrated to other site and formed new cell clone. (f) Suspending cell aggregates. (g) An oocyte-like cell. (h) A blastocyst -like structure attaching to a cell aggregate. (i) PCR analysis showed that the expression of Oct4, Sox2, Nanog, Stellar, Ifitm3, Nanos3, Dazl, and Vasa in cultures at day 7 and day16 after single-cell culture. Immunocytochemistry showed that Dazl positive cells (j, k) and Vasa positive cells (l) appeared in cultures at day 16. Scale bars = 10 μm in (d, f-h), 20 μm in (a-c), 25 μm in (j-l), 40 μm in (e).
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f6: Generation of germline-like cells from a single cell.(a) Morphology of a single L929 cell in culture. (b) The isolated L929 cells showed fibroblast-shaped cells at about day 2. (c) Morphology of the single clone at day 5. (d) Round cells appeared above fibroblast-shaped cells in single-cell culture at day 8. (e-h) The cultures were observed at day 16. (e) Round-shaped cells migrated to other site and formed new cell clone. (f) Suspending cell aggregates. (g) An oocyte-like cell. (h) A blastocyst -like structure attaching to a cell aggregate. (i) PCR analysis showed that the expression of Oct4, Sox2, Nanog, Stellar, Ifitm3, Nanos3, Dazl, and Vasa in cultures at day 7 and day16 after single-cell culture. Immunocytochemistry showed that Dazl positive cells (j, k) and Vasa positive cells (l) appeared in cultures at day 16. Scale bars = 10 μm in (d, f-h), 20 μm in (a-c), 25 μm in (j-l), 40 μm in (e).

Mentions: There are two possible explanations for the spontaneous germline potential of L929 cells. One possibility is that germ cells or ES/EC cells are present among the fibroblast cells. Alternatively, a subpopulation of differentiated cells might reacquire germline potential, due to carcinogenic mutations or epigenetic modifications. Single clone analysis was performed to observe the process of germline-like cell formation from a single differentiated cell. Most of the single cells differentiated and expanded at the beginning (Figure 6a–c). At approximately day 8, round cells appeared in the single-cell cultures (Figure 6d). Upon further culture, the round cells proliferated and formed cell clusters. At day 16, suspending cell aggregates (Figure 6f) and individual germline-like cells (not shown) formed. A subpopulation of round cells migrated to another site and formed new clones (Figure 6e), similar to germline migration. Oocyte-like cells (Figure 6g) and embryo-like structures (Figure 6h) could be observed at day 16 in cultures. The results of the RT-PCR showed that Oct4, Sox2, Nanog, Stellar, Nanos3, DAZL, and Vasa were almost undetectable at day7 but were detected at day 16 (Figure 6i) in cultures. The level of Ifitm3 mRNA was low at day7 but higher at day 16 (Figure 6i) in cultures. Immunohistochemistry staining showed that DAZL (Figure 6j, k) and Vasa expressed in germline-like cells, suggesting their similarity to germ cells.


Spontaneous generation of germline characteristics in mouse fibrosarcoma cells.

Ma Z, Hu Y, Jiang G, Hou J, Liu R, Lu Y, Liu C - Sci Rep (2012)

Generation of germline-like cells from a single cell.(a) Morphology of a single L929 cell in culture. (b) The isolated L929 cells showed fibroblast-shaped cells at about day 2. (c) Morphology of the single clone at day 5. (d) Round cells appeared above fibroblast-shaped cells in single-cell culture at day 8. (e-h) The cultures were observed at day 16. (e) Round-shaped cells migrated to other site and formed new cell clone. (f) Suspending cell aggregates. (g) An oocyte-like cell. (h) A blastocyst -like structure attaching to a cell aggregate. (i) PCR analysis showed that the expression of Oct4, Sox2, Nanog, Stellar, Ifitm3, Nanos3, Dazl, and Vasa in cultures at day 7 and day16 after single-cell culture. Immunocytochemistry showed that Dazl positive cells (j, k) and Vasa positive cells (l) appeared in cultures at day 16. Scale bars = 10 μm in (d, f-h), 20 μm in (a-c), 25 μm in (j-l), 40 μm in (e).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3473365&req=5

f6: Generation of germline-like cells from a single cell.(a) Morphology of a single L929 cell in culture. (b) The isolated L929 cells showed fibroblast-shaped cells at about day 2. (c) Morphology of the single clone at day 5. (d) Round cells appeared above fibroblast-shaped cells in single-cell culture at day 8. (e-h) The cultures were observed at day 16. (e) Round-shaped cells migrated to other site and formed new cell clone. (f) Suspending cell aggregates. (g) An oocyte-like cell. (h) A blastocyst -like structure attaching to a cell aggregate. (i) PCR analysis showed that the expression of Oct4, Sox2, Nanog, Stellar, Ifitm3, Nanos3, Dazl, and Vasa in cultures at day 7 and day16 after single-cell culture. Immunocytochemistry showed that Dazl positive cells (j, k) and Vasa positive cells (l) appeared in cultures at day 16. Scale bars = 10 μm in (d, f-h), 20 μm in (a-c), 25 μm in (j-l), 40 μm in (e).
Mentions: There are two possible explanations for the spontaneous germline potential of L929 cells. One possibility is that germ cells or ES/EC cells are present among the fibroblast cells. Alternatively, a subpopulation of differentiated cells might reacquire germline potential, due to carcinogenic mutations or epigenetic modifications. Single clone analysis was performed to observe the process of germline-like cell formation from a single differentiated cell. Most of the single cells differentiated and expanded at the beginning (Figure 6a–c). At approximately day 8, round cells appeared in the single-cell cultures (Figure 6d). Upon further culture, the round cells proliferated and formed cell clusters. At day 16, suspending cell aggregates (Figure 6f) and individual germline-like cells (not shown) formed. A subpopulation of round cells migrated to another site and formed new clones (Figure 6e), similar to germline migration. Oocyte-like cells (Figure 6g) and embryo-like structures (Figure 6h) could be observed at day 16 in cultures. The results of the RT-PCR showed that Oct4, Sox2, Nanog, Stellar, Nanos3, DAZL, and Vasa were almost undetectable at day7 but were detected at day 16 (Figure 6i) in cultures. The level of Ifitm3 mRNA was low at day7 but higher at day 16 (Figure 6i) in cultures. Immunohistochemistry staining showed that DAZL (Figure 6j, k) and Vasa expressed in germline-like cells, suggesting their similarity to germ cells.

Bottom Line: In this study, we further showed that cells functioning as germline could be present in mouse fibrosarcoma cells (L929 cell line).Early germline-like cells spontaneously appeared in L929 cells and further differentiated into oocyte-like cells.These germline-like cells can, in turn, develop into blastocyst-like structures in vitro and cause teratocarcinomas in vivo, which is consistent with natural germ cells in function.

View Article: PubMed Central - PubMed

Affiliation: Department of Labratory Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai 200040, China.

ABSTRACT
Germline/embryonic-specific genes have been found to be activated in somatic tumors. In this study, we further showed that cells functioning as germline could be present in mouse fibrosarcoma cells (L929 cell line). Early germline-like cells spontaneously appeared in L929 cells and further differentiated into oocyte-like cells. These germline-like cells can, in turn, develop into blastocyst-like structures in vitro and cause teratocarcinomas in vivo, which is consistent with natural germ cells in function. Generation of germline-like cells from somatic tumors might provide a novel way to understand why somatic cancer cells have strong features of embryonic/germline development. It is thought that the germline traits of tumors are associated with the central characteristics of malignancy, such as immortalization, invasion, migration and immune evasion. Therefore, germline-like cells in tumors might provide potential targets to tumor biology, diagnosis and therapy.

Show MeSH
Related in: MedlinePlus