Synthetic extracellular matrices and astrocytes provide a supportive microenvironment for the cultivation and investigation of primary pediatric gliomas

Abstract

Background Pediatric gliomas comprise a diverse set of brain tumor entities that have substantial long-term ramifications for patient survival and quality of life. However, the study of these tumors is currently limited due to a lack of authentic models. Additionally, many aspects of pediatric brain tumor biology, such as tumor cell invasiveness, have been difficult to study with currently available tools. To address these issues, we developed a synthetic extracellular matrix (sECM)-based culture system to grow and study primary pediatric brain tumor cells. Methods We developed a brain-like sECM material as a supportive scaffold for the culture of primary, patient-derived pediatric glioma cells and established patient-derived cell lines. Primary juvenile brainstem-derived murine astrocytes were used as a feeder layer to support the growth of primary human tumor cells. Results We found that our culture system facilitated the proliferation of various primary pediatric brain tumors, including low-grade gliomas, and enabled ex vivo testing of investigational therapeutics. Additionally, we found that tuning this sECM material allowed us to assess high-grade pediatric glioma cell invasion and evaluate therapeutic interventions targeting invasive behavior. Conclusion Our sECM culture platform provides a multipurpose tool for pediatric brain tumor researchers that enables both a wide breadth of biological assays and the cultivation of diverse tumor types.

Generation of patient-derived pediatric pilocytic astrocytoma in-vitro models using SV40 large T: evaluation of a modeling workflow

“…SV40-TAg PA models are less cost effective compared to e.g. cultures in synthetic extracellular matrices [ 8 ]. Taken together, they are excellent and unique tools for standardized studies of tumor cell intrinsic mechanisms and preclinical drug testing to uncover new therapeutic approaches, such as rational combination treatments [ 12 ] or senolytic BH3-mimetics [ 17 ].…”

“…Loss of NF1 , absence of atypical alterations (such as ATRX -loss or CDKN2A -loss) and genetic stability over time support JHH_NF1_PA1 to be a true PA model. Other approaches to culture patient-derived pLGG cells beyond classical culture methods include mouse brain-slice overlay cultures [ 7 ] or synthetic extracellular matrices with astrocytes [ 8 ]. These latter techniques amend the portfolio of preclinical pLGG models but are not ideally suitable for large-scale comprehensive preclinical testing.…”

Pediatric low-grade glioma models: advances and ongoing challenges

“…(2022) developed an ex vivo culture system that utilized synthetic extracellular matrices (sECMs) to partially mimic properties of extracellular matrix in vivo . This culture method promoted proliferation and enabled propagation of several patient-derived pLGG cells for up to 1 month ex vivo ( 57 ). Continuous efforts are made to generate more pLGG patient-derived cell lines that can be cultured long enough for preclinical testing and still resemble the original tumors, but these studies have made major advancements in the field.…”

“…For example, since there is such strong dependence of pLGG formation in vivo on microenvironment, certain factors could be added into the cultures such as cytokines or tumor cells could be co-cultured with other cell types to provide more supportive microenvironment. The relatively successful sECM method to culture pLGG cells demonstrates that elements in the microenvironment will be necessary to generate viable in vitro pLGG models ( 57 ). There is also a need to develop more sophisticated technologies for genetic manipulation of tissues and cells.…”

“…The overexpression of hTERT (human telomerase reverse transcriptase, the catalytic subunit of telomerase) was proposed to counteract OIS in pLGG cell lines so they regain their proliferative potential for long-term cultures [ (55), Figure 2 56), Figure 2)]. They found that these culture conditions led to reversible blockage of senescence, increased proliferation, and allowed these cell lines to propagate longer in culture while still maintaining signature genetic changes of the original tumors (56). This method was tried on multiple pLGG cell lines including 4 PAs, 2 gangliogliomas, 2 anaplastic gliomas, 1 anaplastic PXA and others.…”