Short-term culture of pediatric brain tumors

Mackillop, William J.; Blundell, John; Steele, Patricia

doi:10.1007/bf00735732

Cited by 4 publications

(4 citation statements)

References 6 publications

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“…28 Cells from human brain tumors have the ability to form multicellular spheroids after an initial enzymatic dissociation of the tumor tissue.~~ Spheroid formation has also been performed in short-term culture of biopsies from brain tumors in children. 18 The present study shows that surgically removed tissue specimens from human gliomas easily form tumor spheroids in agar overlay culture. Their morphological features and the individual growth of different spheroids have been studied, from the same patient as well as from different patients.…”

mentioning

confidence: 52%

Multicellular tumor spheroids from human gliomas maintained in organ culture

Bjerkvig¹,

Tønnesen²,

Lærum³

et al. 1990

Journal of Neurosurgery

177

160

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Tumor tissue from seven human gliomas was maintained in long-term agar overlay culture as multicellular organotypic spheroids. Light microscopic and ultrastructural observation of the spheroids displayed morphological features similar to those of the original tumor tissue in vivo; in this respect they were different from spheroids obtained from permanent cell lines. The spheroids contained preserved vessels, connective tissue, and macrophages, revealing a close resemblance to the conditions in the original tumor. Flow cytometric deoxyribonucleic acid measurements of cells from the tumor spheroids and from biopsy material obtained directly from the operation revealed the same ploidy and the same amount of proliferating cells in the spheroids as in the original tumor. Fluorescence microscopy using bromodeoxyuridine (BUdR) incorporation and anti-BUdR monoclonal antibody confirmed the proliferative potential of tumor cells in the spheroids. Diameter measurements showed that the size of the spheroids from two of the tumors increased over time while in three other cases it decreased. Spheroids from the remaining two tumors showed no change in size, even after 80 days in culture. These growth data and the relatively high number of proliferating cells, as measured by flow cytometry, indicate that the degree of cell proliferation and cell loss from the spheroids are closely linked, as is the case for tumors in vivo. The culture system presented provides a valuable alternative to propagation of human tumors in animals.

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mentioning

confidence: 52%

Multicellular tumor spheroids from human gliomas maintained in organ culture

Bjerkvig¹,

Tønnesen²,

Lærum³

et al. 1990

Journal of Neurosurgery

177

160

View full text Add to dashboard Cite

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“…These cells have the ability to self-renew, proliferate and differentiate, and uniquely maintain the tumor growth. Although qualitative reports of sphere formation in cultured brain tumor cells exist (Mackillop et al, 1985) and stem-like cells have been observed in brain tumor cultures (Ignatova et al, 2002), a prospectively isolated population of cancer stem cells from brain tumors had not been described until recently (Singh et al, 2003). The existence of cancer stem cells, first proven in the context of acute myeloid leukemia (AML), required sorting by surface markers to distinguish leukemic stem cells from the remaining AML cells, which had limited proliferative potential (Blair et al, 1997;Bonnet and Dick, 1997).…”

Section: Prospective Identification Of a Cancer Stem Cell From Human mentioning

confidence: 99%

Cancer stem cells in nervous system tumors

et al. 2004

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Most current research on human brain tumors is focused on the molecular and cellular analysis of the bulk tumor mass. However, evidence in leukemia and more recently in solid tumors such as breast cancer suggests that the tumor cell population is heterogeneous with respect to proliferation and differentiation. Recently, several groups have described the existence of a cancer stem cell population in human brain tumors of different phenotypes from both children and adults. The finding of brain tumor stem cells (BTSCs) has been made by applying the principles for cell culture and analysis of normal neural stem cells (NSCs) to brain tumor cell populations and by identification of cell surface markers that allow for isolation of distinct tumor cell populations that can then be studied in vitro and in vivo. A population of brain tumor cells can be enriched for BTSCs by cell sorting of dissociated suspensions of tumor cells for the NSC marker CD133. These CD133 þ cells, which also expressed the NSC marker nestin, but not differentiated neural lineage markers, represent a minority fraction of the entire brain tumor cell population, and exclusively generate clonal tumor spheres in suspension culture and exhibit increased self-renewal capacity. BTSCs can be induced to differentiate in vitro into tumor cells that phenotypically resembled the tumor from the patient. Here, we discuss the evidence for and implications of the discovery of a cancer stem cell in human brain tumors. The identification of a BTSC provides a powerful tool to investigate the tumorigenic process in the central nervous system and to develop therapies targeted to the BTSC. Specific genetic and molecular analyses of the BTSC will further our understanding of the mechanisms of brain tumor growth, reinforcing parallels between normal neurogenesis and brain tumorigenesis.

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“…They are believed to be a better model than monolayer cultures due to a three-dimensional structure with more in vivo-like intercellular contacts. This model was later on further improved by deriving spheroids from single cells obtained from dissociated primary glioblastoma tissue (Mackillop et al, 1985). In order to obtain an even more in vivo-like model the primary organotypic spheroids were introduced .…”

Section: Establishment and Development Of The Spheroid Modelmentioning

confidence: 99%

“…However, the three-dimensional aspect came into focus in the 1970's, where scientists started to grow tumor cells from cell lines as multicellular spheroids (Yuhas et al, 1977). Over the years the spheroid model has been improved by deriving spheroids from cells obtained from dissociated primary glioblastoma tissue (Mackillop et al, 1985) as well as by using organotypic primary spheroids derived from small tumor fragments . In general, most in vitro studies are performed with cells cultured in conventional serumcontaining medium.…”

Section: Introductionmentioning

confidence: 99%