Limits...
Ultrasonic Surgical Aspirate is a Reliable Source For Culturing Glioblastoma Stem Cells

View Article: PubMed Central - PubMed

ABSTRACT

Glioma stem cells (GSCs) are thought to be the source of tumor growth and therapy resistance. To understand the biology of GSCs, and target these tumors therapeutically, we need robust strategies for in vitro expansion of primary GSCs. To date, tumor core biopsies have been the main established source of GSCs. Since these samples are used for diagnostic purposes, the available tissue for cell culture and therapeutic targeting can be limited. In addition, a core biopsy is usually taken from one part of the tumor, thus would be unlikely to represent intra-tumor heterogeneity. To overcome these problems, tissue fragments from all over the tumor can be collected using an ultrasonic aspirator during surgery, thus assembling a “global tumor biopsy”. Usually, this ultrasonic aspirate (UA) sample is considered as biological waste after operations. Here, we show that UA samples offer a large and reliable source of live cells. Similar to core biopsies, UA samples enriched for GSCs that differentiated into neural lineages, showed inter-individual variation of GSC markers, and induced tumors. Molecular profiling showed that UA samples cover tumor heterogeneity better than core biopsies. These results suggest that UA samples can be used to establish large scale cultures for therapeutic applications.

No MeSH data available.


Related in: MedlinePlus

UA cells have multilineage neural differentiation potential similar to tumor core-derived cells.Staining for neuronal, glial and oligodendrocytes markers after two weeks of differentiation for UA cells (first column) and tumor core-derived cells (third column) show that differentiated cells from both samples, UA and Core, express higher levels of GFAP, BIII and down regulate the expression of immature neural stem cell markers such as Nestin and DCx than undifferentiated control cells. The same laser intensity and confocal settings were used for the comparison. Scale bar: 20 μM.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5015049&req=5

f5: UA cells have multilineage neural differentiation potential similar to tumor core-derived cells.Staining for neuronal, glial and oligodendrocytes markers after two weeks of differentiation for UA cells (first column) and tumor core-derived cells (third column) show that differentiated cells from both samples, UA and Core, express higher levels of GFAP, BIII and down regulate the expression of immature neural stem cell markers such as Nestin and DCx than undifferentiated control cells. The same laser intensity and confocal settings were used for the comparison. Scale bar: 20 μM.

Mentions: To examine the neural differentiation potential of UA cells, we subjected them to a neural differentiation protocol for two weeks in the presence of serum, B27 and vitamin A and the absence of growth factors. Under neural differentiation conditions, cells from sphere cultures of both UA and tumor core samples gave rise to cells that expressed markers of mature neurons and glial cells (MAP2, β-TubIII and GFAP), but not oligdendrocytes (O4). However, the differentiated cells did not lose the expression of immature neural/neural stem cell markers (Musashi, DCX and Nestin), but they had reduced expression level of DCX and Nestin compared to undifferentiated cells (Fig. 5). The same laser intensity and confocal settings were used for UA and the matched core samples compared to their undifferentiated cells. This assay was done on samples from five patients. Thus, sphere culture cells from both UA and tumor core biopsies have similar multilineage neural differentiation capabilities.


Ultrasonic Surgical Aspirate is a Reliable Source For Culturing Glioblastoma Stem Cells
UA cells have multilineage neural differentiation potential similar to tumor core-derived cells.Staining for neuronal, glial and oligodendrocytes markers after two weeks of differentiation for UA cells (first column) and tumor core-derived cells (third column) show that differentiated cells from both samples, UA and Core, express higher levels of GFAP, BIII and down regulate the expression of immature neural stem cell markers such as Nestin and DCx than undifferentiated control cells. The same laser intensity and confocal settings were used for the comparison. Scale bar: 20 μM.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015049&req=5

f5: UA cells have multilineage neural differentiation potential similar to tumor core-derived cells.Staining for neuronal, glial and oligodendrocytes markers after two weeks of differentiation for UA cells (first column) and tumor core-derived cells (third column) show that differentiated cells from both samples, UA and Core, express higher levels of GFAP, BIII and down regulate the expression of immature neural stem cell markers such as Nestin and DCx than undifferentiated control cells. The same laser intensity and confocal settings were used for the comparison. Scale bar: 20 μM.
Mentions: To examine the neural differentiation potential of UA cells, we subjected them to a neural differentiation protocol for two weeks in the presence of serum, B27 and vitamin A and the absence of growth factors. Under neural differentiation conditions, cells from sphere cultures of both UA and tumor core samples gave rise to cells that expressed markers of mature neurons and glial cells (MAP2, β-TubIII and GFAP), but not oligdendrocytes (O4). However, the differentiated cells did not lose the expression of immature neural/neural stem cell markers (Musashi, DCX and Nestin), but they had reduced expression level of DCX and Nestin compared to undifferentiated cells (Fig. 5). The same laser intensity and confocal settings were used for UA and the matched core samples compared to their undifferentiated cells. This assay was done on samples from five patients. Thus, sphere culture cells from both UA and tumor core biopsies have similar multilineage neural differentiation capabilities.

View Article: PubMed Central - PubMed

ABSTRACT

Glioma stem cells (GSCs) are thought to be the source of tumor growth and therapy resistance. To understand the biology of GSCs, and target these tumors therapeutically, we need robust strategies for in vitro expansion of primary GSCs. To date, tumor core biopsies have been the main established source of GSCs. Since these samples are used for diagnostic purposes, the available tissue for cell culture and therapeutic targeting can be limited. In addition, a core biopsy is usually taken from one part of the tumor, thus would be unlikely to represent intra-tumor heterogeneity. To overcome these problems, tissue fragments from all over the tumor can be collected using an ultrasonic aspirator during surgery, thus assembling a “global tumor biopsy”. Usually, this ultrasonic aspirate (UA) sample is considered as biological waste after operations. Here, we show that UA samples offer a large and reliable source of live cells. Similar to core biopsies, UA samples enriched for GSCs that differentiated into neural lineages, showed inter-individual variation of GSC markers, and induced tumors. Molecular profiling showed that UA samples cover tumor heterogeneity better than core biopsies. These results suggest that UA samples can be used to establish large scale cultures for therapeutic applications.

No MeSH data available.


Related in: MedlinePlus