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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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Further differentiation of early PGCs.(a) Suspending cell aggregates. (b) Suspending individual germline-like cells. (c) Appearance of bigger cells. (d) A bigger germline-like cells in high magnification. Expression of c-kit protein in a subpopulation of germline-like cells. Immunocytochemistry showed that DAZL was detected in germline-like cell aggregates (e), suspending cell aggregates (f), and individual germline-like cells with different diameter (g). Immunocytochemistry showed that Vasa were detected in cell aggregates (h) and individual germline-like cells (i). (j) Flow cytometry analysis showed the ratio of Vasa+ cells were in cultures at day 25. Scale bars = 10 μm in (a-d), 25 μm in (e-i).
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f2: Further differentiation of early PGCs.(a) Suspending cell aggregates. (b) Suspending individual germline-like cells. (c) Appearance of bigger cells. (d) A bigger germline-like cells in high magnification. Expression of c-kit protein in a subpopulation of germline-like cells. Immunocytochemistry showed that DAZL was detected in germline-like cell aggregates (e), suspending cell aggregates (f), and individual germline-like cells with different diameter (g). Immunocytochemistry showed that Vasa were detected in cell aggregates (h) and individual germline-like cells (i). (j) Flow cytometry analysis showed the ratio of Vasa+ cells were in cultures at day 25. Scale bars = 10 μm in (a-d), 25 μm in (e-i).

Mentions: At 12 days after re-plate, a subpopulation (approximately 10%) of cell aggregates and individual germ-like cells began to detach from the plate and became motile (Figure 2a, b), suggesting that these cells loss cell-cell contact which might be similar to migratory/postmigratory germline cells26. At about day 20, approximately 70% germline-like cells and cell aggregates suspended in media. At day 12, approximately 5% of round-shaped cells increased gradually in diameter (Figure 2c, d), further indicating that the germline-like cells could undergo further development. DAZL is known to be expressed in germ cells and is required for the development of PGCs and for their differentiation and maturation13. Expression of the RNA helicase enzyme Vasa begins in post-migratory PCGs and lasts until the postmeiotic stage of oocytes26. Therefore, Vasa is useful indicator of the presence of post-migratory PGCs2526. Immunocytochemistry staining showed that both DAZL (Figure 2e–g) and Vasa (Figure 2h, i) are expressed in a subpopulation of individual germline-like cells and cell aggregates. The appearance of Vasa+ germline-like cells suggested that we were observing postmigratory PGC-like cells2526. The results of the RT-PCR experiments further showed that DAZL and Vasa were expressed in the cultures (Figure 1l). Flow cytometry analysis showed that the ratio of Vasa+ cells were approximately 22.96±1.45% (n = 3) of the total cell population at day 20 (Figure 2j).


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)

Further differentiation of early PGCs.(a) Suspending cell aggregates. (b) Suspending individual germline-like cells. (c) Appearance of bigger cells. (d) A bigger germline-like cells in high magnification. Expression of c-kit protein in a subpopulation of germline-like cells. Immunocytochemistry showed that DAZL was detected in germline-like cell aggregates (e), suspending cell aggregates (f), and individual germline-like cells with different diameter (g). Immunocytochemistry showed that Vasa were detected in cell aggregates (h) and individual germline-like cells (i). (j) Flow cytometry analysis showed the ratio of Vasa+ cells were in cultures at day 25. Scale bars = 10 μm in (a-d), 25 μm in (e-i).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Further differentiation of early PGCs.(a) Suspending cell aggregates. (b) Suspending individual germline-like cells. (c) Appearance of bigger cells. (d) A bigger germline-like cells in high magnification. Expression of c-kit protein in a subpopulation of germline-like cells. Immunocytochemistry showed that DAZL was detected in germline-like cell aggregates (e), suspending cell aggregates (f), and individual germline-like cells with different diameter (g). Immunocytochemistry showed that Vasa were detected in cell aggregates (h) and individual germline-like cells (i). (j) Flow cytometry analysis showed the ratio of Vasa+ cells were in cultures at day 25. Scale bars = 10 μm in (a-d), 25 μm in (e-i).
Mentions: At 12 days after re-plate, a subpopulation (approximately 10%) of cell aggregates and individual germ-like cells began to detach from the plate and became motile (Figure 2a, b), suggesting that these cells loss cell-cell contact which might be similar to migratory/postmigratory germline cells26. At about day 20, approximately 70% germline-like cells and cell aggregates suspended in media. At day 12, approximately 5% of round-shaped cells increased gradually in diameter (Figure 2c, d), further indicating that the germline-like cells could undergo further development. DAZL is known to be expressed in germ cells and is required for the development of PGCs and for their differentiation and maturation13. Expression of the RNA helicase enzyme Vasa begins in post-migratory PCGs and lasts until the postmeiotic stage of oocytes26. Therefore, Vasa is useful indicator of the presence of post-migratory PGCs2526. Immunocytochemistry staining showed that both DAZL (Figure 2e–g) and Vasa (Figure 2h, i) are expressed in a subpopulation of individual germline-like cells and cell aggregates. The appearance of Vasa+ germline-like cells suggested that we were observing postmigratory PGC-like cells2526. The results of the RT-PCR experiments further showed that DAZL and Vasa were expressed in the cultures (Figure 1l). Flow cytometry analysis showed that the ratio of Vasa+ cells were approximately 22.96±1.45% (n = 3) of the total cell population at day 20 (Figure 2j).

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