Isolation and characterization of stem cell-like precursor cells from primary human anaplastic oligoastrocytoma

Abstract

A small population of stem cell-like precursors in solid tumors are linked to histological composition, progression, angiogenesis, metastasis, recurrence and drug resistance of a variety of malignant tumors. Oligoastrocytoma is the most common brain mixed glioma composed of mixed cells of oligodendroglial and astrocytic phenotypes. Identification and characterization of stem cell-like precursors in oligoastrocytoma may shed light on the oncogenesis of this unique type of tumor and assist in the design of novel therapeutic strategy. Here, tumor stem cell-like precursors were identified from primary human anaplastic oligoastrocytomas by labeling of the tumor sections with nestin and CD133. Tumor cells were cultured in vitro in stem cell medium with growth factors and the capacity of the surviving stem cell-like precursors to form tumor spheres was tested. The tumor spheres were further injected subcutaneously into nude mice to observe the contribution of stem cell-like precursors to histological composition and tumor progression. We found that primary human oligoastrocytoma tissues contained nestin þ /CD133 þ stem cell-like precursors. These cells differentiated into tumor cells with both oligodendroglial and astrocytic characteristics and formed tumor spheres in vitro, which upon implantation in nude mice, grew into tumor nodules containing nestin þ /CD133 þ cells at levels higher than in the primary tumor tissues. This study revealed for the first time that anaplastic human oligoastrocytomas contained stem cell-like precursors, which exhibit neural stem cell properties with tumorigenicity. These stem cell-like precursors may be responsible for the oligodendroglial and astrocytic components of human oligoastrocytoma and should be considered as therapeutic targets.

A simplified and modified procedure to culture brain glioma stem cells from clinical specimens

“…As the origins of glioma cells and the critical factors, BGSCs determine glioma propagation, progression and therapeutic resistance. Subsequently, BGSCs with a tumor-initiating ability have been isolated from GBMs, medulloblastomas, ependymomas and anaplastic oligoastrocytomas, and certain biological characteristics have been investigated (4,5,8,11,15,16). These findings suggest a hierarchical model in which glioma arises from BGSCs and progresses through mechanisms similar to a developmental process.…”

“…Tumor-sphere cultures were performed according to reported procedures (4)(5)(6)8), with minor modifications. Following resection, the tumor tissues were washed, minced in phosphatebuffered saline (PBS) and subjected to enzymatic dissociation.…”

High expression of leptin receptor leads to temozolomide resistance with exhibiting stem/progenitor cell features in gliobalastoma

“…The purity of the ObR cells was analyzed using a similar protocol, and the cells were maintained in stem cell medium as previously described. 38 The cells' apoptosis was detected by using Annexin V-FITC Apoptosis Detection Kit (BD Biosciences) according to the manufacturer's instructions. Briefly speaking, the pretreated cells were harvested and stained with PI and Annexin V-FITC, the apoptosis rate was assayed using FACS at 488 nm.…”

CD133 Expression and Identification of CD133/nestin Positive Cells in Rhabdomyosarcomas and Rhabdomyosarcoma Cell Lines

“…This was experimentally proven in glioblastoma multiforme [19,27] and in melanoma [23]. Furthermore, the coexpression of CD133 and nestin was also shown in medulloblastomas [33], pilocytic astrocytomas [33], oligoastrocytomas [45], and in osteosarcomas [42].…”