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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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Formation and marker expression in PGC-like cells.(a) Phase contrast images of L929 cells in culture. (b) Small, round cells appeared above the L929 cells at day 7 of culture. (c) Small, round cells in high magnification. (d) Formation of cell aggregates. (e) A bigger cell aggregate. (f) A few germ-like cells positive for AP. Expression of Vim (g) and fibrin (h) in fibroblast-like cells. Oct4 (i), Sox2 (j), and SSEA1 (k) were detected in cultures with germ-like cells. (l) The results of the RT-PCR analysis, showing the genes that were related to germ cell formation expressed in cultures at day 12. The DNA size makers (M) are indicated in the first lane. Normal subcutaneous areolar and adipose tissue (Nt) and ovary (Ov.) was used as a control. (m) Flow cytometry analysis showed that the ratio of SSEA1+ cells were 0.5±0.72% (n = 3) and 30.17±3.98% (n = 3) of in cultures at day 3 and day 12 respectively (P<0.05). Scale bars = 20 μm.
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f1: Formation and marker expression in PGC-like cells.(a) Phase contrast images of L929 cells in culture. (b) Small, round cells appeared above the L929 cells at day 7 of culture. (c) Small, round cells in high magnification. (d) Formation of cell aggregates. (e) A bigger cell aggregate. (f) A few germ-like cells positive for AP. Expression of Vim (g) and fibrin (h) in fibroblast-like cells. Oct4 (i), Sox2 (j), and SSEA1 (k) were detected in cultures with germ-like cells. (l) The results of the RT-PCR analysis, showing the genes that were related to germ cell formation expressed in cultures at day 12. The DNA size makers (M) are indicated in the first lane. Normal subcutaneous areolar and adipose tissue (Nt) and ovary (Ov.) was used as a control. (m) Flow cytometry analysis showed that the ratio of SSEA1+ cells were 0.5±0.72% (n = 3) and 30.17±3.98% (n = 3) of in cultures at day 3 and day 12 respectively (P<0.05). Scale bars = 20 μm.

Mentions: The L929 cells exhibited adherent growth and heterogeneous in morphology, including spindle-like, epithelial-like, stellate, and round shape (Figure 1a). The L929 cells expressed Vim (Figure 1g) and Fib (Figure 1h). To facilitate the study of germ cell formation, the cells were continuously cultured for 2–4 weeks without subculture such that they reached full confluence and high density. Approximately 7 days later after re-plate, the adherent differentiating cells proliferated to form a confluent layer of cells, and the small round-shaped cells with a high nucleus-to-cytoplasm ratio, resembling primordial germ cells (PGCs), grew above the confluent layer in culture (Figure 1b, c). The small, round-shaped cells were approximately 5–8 μm in diameter at the beginning. After about 10 days of culture, these small, round-shaped cells formed cellular aggregates (Figure 1d, e) that were similar to the clusters formed by PGCs in vivo25. The shape of the cellular aggregates was different from the granulated colonies of EC/ES cells. The results suggested that germline-like cells might appear in cultures.


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)

Formation and marker expression in PGC-like cells.(a) Phase contrast images of L929 cells in culture. (b) Small, round cells appeared above the L929 cells at day 7 of culture. (c) Small, round cells in high magnification. (d) Formation of cell aggregates. (e) A bigger cell aggregate. (f) A few germ-like cells positive for AP. Expression of Vim (g) and fibrin (h) in fibroblast-like cells. Oct4 (i), Sox2 (j), and SSEA1 (k) were detected in cultures with germ-like cells. (l) The results of the RT-PCR analysis, showing the genes that were related to germ cell formation expressed in cultures at day 12. The DNA size makers (M) are indicated in the first lane. Normal subcutaneous areolar and adipose tissue (Nt) and ovary (Ov.) was used as a control. (m) Flow cytometry analysis showed that the ratio of SSEA1+ cells were 0.5±0.72% (n = 3) and 30.17±3.98% (n = 3) of in cultures at day 3 and day 12 respectively (P<0.05). Scale bars = 20 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Formation and marker expression in PGC-like cells.(a) Phase contrast images of L929 cells in culture. (b) Small, round cells appeared above the L929 cells at day 7 of culture. (c) Small, round cells in high magnification. (d) Formation of cell aggregates. (e) A bigger cell aggregate. (f) A few germ-like cells positive for AP. Expression of Vim (g) and fibrin (h) in fibroblast-like cells. Oct4 (i), Sox2 (j), and SSEA1 (k) were detected in cultures with germ-like cells. (l) The results of the RT-PCR analysis, showing the genes that were related to germ cell formation expressed in cultures at day 12. The DNA size makers (M) are indicated in the first lane. Normal subcutaneous areolar and adipose tissue (Nt) and ovary (Ov.) was used as a control. (m) Flow cytometry analysis showed that the ratio of SSEA1+ cells were 0.5±0.72% (n = 3) and 30.17±3.98% (n = 3) of in cultures at day 3 and day 12 respectively (P<0.05). Scale bars = 20 μm.
Mentions: The L929 cells exhibited adherent growth and heterogeneous in morphology, including spindle-like, epithelial-like, stellate, and round shape (Figure 1a). The L929 cells expressed Vim (Figure 1g) and Fib (Figure 1h). To facilitate the study of germ cell formation, the cells were continuously cultured for 2–4 weeks without subculture such that they reached full confluence and high density. Approximately 7 days later after re-plate, the adherent differentiating cells proliferated to form a confluent layer of cells, and the small round-shaped cells with a high nucleus-to-cytoplasm ratio, resembling primordial germ cells (PGCs), grew above the confluent layer in culture (Figure 1b, c). The small, round-shaped cells were approximately 5–8 μm in diameter at the beginning. After about 10 days of culture, these small, round-shaped cells formed cellular aggregates (Figure 1d, e) that were similar to the clusters formed by PGCs in vivo25. The shape of the cellular aggregates was different from the granulated colonies of EC/ES cells. The results suggested that germline-like cells might appear in cultures.

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