Limits...
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.

Show MeSH

Related in: MedlinePlus

Formation of oocyte-like cells.(a) Phase contrast images of an oocyte-like cell with a GV-like structure. (b) A bigger oocyte-like cells. (c) Large oocyte-like cells lacking a zona pellucida membrane. (d-f) Immunocytochemistry showing the expression of DAZL protein in a larger cell. (d) DAZL staining. (e) DAPI staining. (f) A merged image of Dazl protein (red) and DAPI (blue). (g-j) Immunocytochemistry showing the expression of Vasa protein in a bigger cell. (g) Phase contrast images of an oocyte-like cell with a GV-like structure. (h) Vasa staining. (i) DAPI staining (nucleolus, arrow). (j) Vasa/DAPI staining. (k-m) Immunocytochemistry showing the expression of Vasa protein in a larger oocyte-like cell. (k) Phase contrast images of the oocyte-like cell. (l) DAPI staining. (m) Vasa/DAPI staining. (n-p) SCP3 protein was detected in a large cell. (n) SCP3 staining. (o) DAPI staining. (p) SCP3 /DAPI staining. (q) The expression of ZP2, ZP3, SCP1, SCP3, and GDF9 mRNA was detected in these big cells (BC) (>20 μm in diameter). (r) The levels of media estradiol were detectable in culture. Blank (B) medium was used as a control. No statistical difference between the two groups (P>0.05). Scale bars = 10 μm in (a-c), 25 μm in (d-p).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3473365&req=5

f3: Formation of oocyte-like cells.(a) Phase contrast images of an oocyte-like cell with a GV-like structure. (b) A bigger oocyte-like cells. (c) Large oocyte-like cells lacking a zona pellucida membrane. (d-f) Immunocytochemistry showing the expression of DAZL protein in a larger cell. (d) DAZL staining. (e) DAPI staining. (f) A merged image of Dazl protein (red) and DAPI (blue). (g-j) Immunocytochemistry showing the expression of Vasa protein in a bigger cell. (g) Phase contrast images of an oocyte-like cell with a GV-like structure. (h) Vasa staining. (i) DAPI staining (nucleolus, arrow). (j) Vasa/DAPI staining. (k-m) Immunocytochemistry showing the expression of Vasa protein in a larger oocyte-like cell. (k) Phase contrast images of the oocyte-like cell. (l) DAPI staining. (m) Vasa/DAPI staining. (n-p) SCP3 protein was detected in a large cell. (n) SCP3 staining. (o) DAPI staining. (p) SCP3 /DAPI staining. (q) The expression of ZP2, ZP3, SCP1, SCP3, and GDF9 mRNA was detected in these big cells (BC) (>20 μm in diameter). (r) The levels of media estradiol were detectable in culture. Blank (B) medium was used as a control. No statistical difference between the two groups (P>0.05). Scale bars = 10 μm in (a-c), 25 μm in (d-p).

Mentions: Upon further culture, round or ovoid-shaped cells were present in cultures (Figure 3a–c), resembling gonocytes or primitive oocytes in morphology. Approximately 98% of the oocyte-like cells were 20–25 μm in diameter (Figure 3a, c). A few oocyte-like cells could reach up to about 40μm (Figure 3c). A zona pellucida-like membrane was unobvious around the oocyte-like cells. The immunocytochemistry results showed that these round, large cells expressed DAZL (Figure 3d–f) and Vasa protein (Figure 3g–m), suggesting their identity as germline cells. To further confirm the identity of the bigger germline-like cells, RT-PCR was performed to detect oocyte-specific genes, including zona pellucida genes (ZP1, ZP2, ZP3) and Growth/differentiation factor 9 (Gdf9). The results of the RT-PCR showed that ZP2, ZP3, and Gdf9 were present in the oocyte-like cells (Figure 3q). However, ZP1 was undetectable in the larger germline-like cells (Figure 3q).


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 of oocyte-like cells.(a) Phase contrast images of an oocyte-like cell with a GV-like structure. (b) A bigger oocyte-like cells. (c) Large oocyte-like cells lacking a zona pellucida membrane. (d-f) Immunocytochemistry showing the expression of DAZL protein in a larger cell. (d) DAZL staining. (e) DAPI staining. (f) A merged image of Dazl protein (red) and DAPI (blue). (g-j) Immunocytochemistry showing the expression of Vasa protein in a bigger cell. (g) Phase contrast images of an oocyte-like cell with a GV-like structure. (h) Vasa staining. (i) DAPI staining (nucleolus, arrow). (j) Vasa/DAPI staining. (k-m) Immunocytochemistry showing the expression of Vasa protein in a larger oocyte-like cell. (k) Phase contrast images of the oocyte-like cell. (l) DAPI staining. (m) Vasa/DAPI staining. (n-p) SCP3 protein was detected in a large cell. (n) SCP3 staining. (o) DAPI staining. (p) SCP3 /DAPI staining. (q) The expression of ZP2, ZP3, SCP1, SCP3, and GDF9 mRNA was detected in these big cells (BC) (>20 μm in diameter). (r) The levels of media estradiol were detectable in culture. Blank (B) medium was used as a control. No statistical difference between the two groups (P>0.05). Scale bars = 10 μm in (a-c), 25 μm in (d-p).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Formation of oocyte-like cells.(a) Phase contrast images of an oocyte-like cell with a GV-like structure. (b) A bigger oocyte-like cells. (c) Large oocyte-like cells lacking a zona pellucida membrane. (d-f) Immunocytochemistry showing the expression of DAZL protein in a larger cell. (d) DAZL staining. (e) DAPI staining. (f) A merged image of Dazl protein (red) and DAPI (blue). (g-j) Immunocytochemistry showing the expression of Vasa protein in a bigger cell. (g) Phase contrast images of an oocyte-like cell with a GV-like structure. (h) Vasa staining. (i) DAPI staining (nucleolus, arrow). (j) Vasa/DAPI staining. (k-m) Immunocytochemistry showing the expression of Vasa protein in a larger oocyte-like cell. (k) Phase contrast images of the oocyte-like cell. (l) DAPI staining. (m) Vasa/DAPI staining. (n-p) SCP3 protein was detected in a large cell. (n) SCP3 staining. (o) DAPI staining. (p) SCP3 /DAPI staining. (q) The expression of ZP2, ZP3, SCP1, SCP3, and GDF9 mRNA was detected in these big cells (BC) (>20 μm in diameter). (r) The levels of media estradiol were detectable in culture. Blank (B) medium was used as a control. No statistical difference between the two groups (P>0.05). Scale bars = 10 μm in (a-c), 25 μm in (d-p).
Mentions: Upon further culture, round or ovoid-shaped cells were present in cultures (Figure 3a–c), resembling gonocytes or primitive oocytes in morphology. Approximately 98% of the oocyte-like cells were 20–25 μm in diameter (Figure 3a, c). A few oocyte-like cells could reach up to about 40μm (Figure 3c). A zona pellucida-like membrane was unobvious around the oocyte-like cells. The immunocytochemistry results showed that these round, large cells expressed DAZL (Figure 3d–f) and Vasa protein (Figure 3g–m), suggesting their identity as germline cells. To further confirm the identity of the bigger germline-like cells, RT-PCR was performed to detect oocyte-specific genes, including zona pellucida genes (ZP1, ZP2, ZP3) and Growth/differentiation factor 9 (Gdf9). The results of the RT-PCR showed that ZP2, ZP3, and Gdf9 were present in the oocyte-like cells (Figure 3q). However, ZP1 was undetectable in the larger germline-like cells (Figure 3q).

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