Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen

Kwak, Jay Jiyong; Shin, Hye Jin; Kim, Se Hoon; Shim, Jae Yong; Lee, Ji Hyun; Huh, Yong Min; Kim, Eui Hyun; Park, Eun Kyung; Chang, Jong Hee; Kim, Sun Ho; Hong, Yong Kil; Kim, Do Kyung; Lee, Su Jae; Kang, Seok‐Gu

doi:10.1007/s00381-013-2201-x

Cited by 14 publications

(16 citation statements)

References 34 publications

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“…Thus, optimal conditions, including rapid transfer of specimens following surgical resection, removal of red blood cells and necrotic material and an appropriate environment for growth are required for the isolation of TSs from fresh surgical specimens [5,7,9,11,12,18,19,23,35]. Our laboratory has an ongoing effort to develop optimal conditions for the isolation of TS [2,3,10,17].…”

Section: Discussionmentioning

confidence: 99%

“…When cultured under appropriate conditions in vitro, this population of tumor cells gives rise to gliomaspheres, referred to more generically as tumorspheres (TSs). TSs have been isolated from various malignant tumors, including breast [4], prostate [5], bone [6], colon [7], kidney [8] and lung [9], as well as brain [3,[10][11][12][13][14]. It was previously reported that the rate of isolation of TSs increase as the World Health Organization (WHO) grades of glioma rise [3].…”

Section: Introductionmentioning

confidence: 99%

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Success of tumorsphere isolation from WHO grade IV gliomas does not correlate with the weight of fresh tumor specimens: an immunohistochemical characterization of tumorsphere differentiation

et al. 2016

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BackgroundA trend of stage-by-stage increase in tumorsphere (TS) formation from glioma samples has been reported. Despite this trend, not all surgical specimens give rise to TSs, even World Health Organization (WHO) grade IV gliomas. Furthermore, it has been reported that differences in overall survival of primary glioblastoma patients depends on the propensity of their tumors to form TSs. However, the weights of fresh specimens vary from one surgical isolate to the next.MethodsAccordingly, we evaluated the relationship between the weights of surgical specimens in WHO grade IV gliomas with the capacity to isolate TSs. Thirty-five fresh WHO grade IV glioma specimens were separated into two groups, based on whether they were positive or negative for TS isolation, and the relationship between TS isolation and weight of surgical specimens was assessed.ResultsWe observed no significant difference in the weights of surgical samples in the two groups, and found that the optimal weight of specimens for TSs isolation was 500 mg.ConclusionThus, contrary to our expectations, the ability to isolate TSs from WHO grade IV glioma specimens was not related to the weight of fresh specimens.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Success of tumorsphere isolation from WHO grade IV gliomas does not correlate with the weight of fresh tumor specimens: an immunohistochemical characterization of tumorsphere differentiation

et al. 2016

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“…Mesenchymal stem-like cells (MSLCs) have been isolated from normal brain [ 6 , 7 ] and Lang et al [ 8 ] presented the isolation of mesenchymal stem cells (MSCs) from glioma for the first time. In addition, tumor MSLCs (tMSLCs) have been isolated from several human brain tumors [ 2 , 9 – 11 ], suggesting that these cells play a role in creating a microenvironment conducive to brain tumor initiation and progression [ 2 , 12 – 14 ].…”

Section: Introductionmentioning

confidence: 99%

Tumor Mesenchymal Stem‐Like Cell as a Prognostic Marker in Primary Glioblastoma

Yoon

Shim

Chang

et al. 2016

Stem Cells International

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The isolation from brain tumors of tumor mesenchymal stem-like cells (tMSLCs) suggests that these cells play a role in creating a microenvironment for tumor initiation and progression. The clinical characteristics of patients with primary glioblastoma (pGBM) positive for tMSLCs have not been determined. This study analyzed samples from 82 patients with pGBM who had undergone tumor removal, pathological diagnosis, and isolation of tMSLC from April 2009 to October 2014. Survival, extent of resection, molecular markers, and tMSLC culture results were statistically evaluated. Median overall survival was 18.6 months, 15.0 months in tMSLC-positive patients and 29.5 months in tMSLC-negative patients (P = 0.014). Multivariate cox regression model showed isolation of tMSLC (OR = 2.5, 95% CI = 1.1~5.6, P = 0.021) showed poor outcome while larger extent of resection (OR = 0.5, 95% CI = 0.2~0.8, P = 0.011) has association with better outcome. The presence of tMSLCs isolated from the specimen of pGBM is associated with the survival of patient.

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“…Neuropathologists diagnosed each surgical specimen according to the WHO classification criteria (19). TSs were isolated from the rPB specimen using a modification of previous methods for TS isolation from human brain cancers (11,13,15,20,21). Briefly, the cell isolation procedure was performed within 60 min of PB removal using a mechanical dissociation method.…”

Section: Methodsmentioning

confidence: 99%

“…Neuro-glial differentiation. The multipotency of rPB TSs was tested by examining neural lineage expression by immunocytochemical staining, as previously described (11,20,26). Briefly, after being seeded onto chamber slides (Lab-Tek II; Nalge Nunc International, Rochester, NY, USA), cells were grown in neural differentiation media containing 10% fetal bovine serum (FBS; Lonza) and 1X B27 supplement (Invitrogen) for up to 14 days.…”

Section: Reverse-transcription Polymerase Chain Reaction (Rt-pcr)mentioning

confidence: 99%

Isolation and characterization of tumorspheres from a recurrent pineoblastoma patient: Feasibility of a patient-derived xenograft

Kwak

Shim

Kim

et al. 2016

International Journal of Oncology

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The existence of tumorspheres (TSs) might confer treatment resistance to pineoblastoma (PB). The existence of PB TSs with cellular immortalization potential has not yet been reported. We developed a procedure for isolating TSs from recurrent PB (rPB) and tested whether their properties made them suitable for use as a patient-derived xenograft (PDX). Immunocytochemical staining, RT-PCR and quantitative real-time PCR showed that, among stemness proteins, CD133, musashi and podoplanin were expressed at elevated levels in rPB TSs, but nestin was not. rPB TSs cultured under neuro-glial differentiation conditions expressed TUBB3, but not GFAP, MBP or NeuN. Unlike glioblastoma TSs, rPB TSs showed no clear evidence of invasion in 3D invasion assay or increased expression of genes associated with epithelial-mesenchymal transition. An orthotopic xenograft showed that tumor xenografts replicated the histopathological features of the patient tumor and expressed similar genome profiles, as determined by short tandem repeat genotyping. These data demonstrate the isolation and the characterization of rPB TSs for the first time. Using an orthotopic xenograft, we showed that rPB TSs could replicate the patient tumor, demonstrating their potential as a PDX for precision medicine.

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Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen

Abstract: We showed that both tumor spheres and MSLCs can be isolated from the same PNET specimen. PNET-MSLCs occupied a niche in the vicinity of the vasculature and could be a source of stroma for PNETs.

Cited by 14 publications

References 34 publications

Success of tumorsphere isolation from WHO grade IV gliomas does not correlate with the weight of fresh tumor specimens: an immunohistochemical characterization of tumorsphere differentiation

Success of tumorsphere isolation from WHO grade IV gliomas does not correlate with the weight of fresh tumor specimens: an immunohistochemical characterization of tumorsphere differentiation

Tumor Mesenchymal Stem‐Like Cell as a Prognostic Marker in Primary Glioblastoma

Isolation and characterization of tumorspheres from a recurrent pineoblastoma patient: Feasibility of a patient-derived xenograft

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