A bioengineering method for modeling alveolar Rhabdomyosarcoma and assessing chemotherapy responses

Autor: David L. Perrin, Ehsan Samiei, Mohammad Esmaeillou, Arya Emami, Saeid Ghavami, Mohsen Akbari, Joseph W. Gordon, Kiarash Roustai Geraylow, Mahboubeh Kavoosi, Evan Stefanek, Milad Ashrafizadeh
Jazyk: angličtina
Rok vydání: 2021
Předmět:
DFS
Disease-free survival
Clinical Biochemistry
Apoptosis
MYOD1
Myogenic muscle differentiation transcription factor 1
Biofabrication
Extracellular matrix
DNA
Deoxyribonucleic acid
0302 clinical medicine
Rhabdomyosarcoma
Medicine
RPMI
Roswell Park Memorial Institute
PDMS
Polydimethylsiloxane
RGD
Arginylglycylaspartic acid
0303 health sciences
IgG
Immunoglobulin G
RMS
Rhabdomyosarcoma
DAPI
4’
6-Diami- dino-2-Phenylindole
Dihydrochloride
EDTA
Ethylenediaminetetraacetic acid
Primary tumor
EM
Engineered model
PAX
Paired box gene
AKT
Protein Kinase B
3. Good health
Medical Laboratory Technology
HEPES
(4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)
030220 oncology & carcinogenesis
Alveolar rhabdomyosarcoma
Protocol Article
Cell death
EthD-1
Ethidium homodimer-1
Programmed cell death
3D
Three-dimensional
Science
dECM
Decellularized extracellular matrix
2D
Two-dimensional
MEK
Mitogen-activated extracellular signal-regulated kinase
FOXO1
Forkhead box protein O1
03 medical and health sciences
BSA
Bovine serum albumin
ECM
Extracellular matrix
In vivo
RT
Room temperature
DMEM
Dulbecco's phosphate buffered saline
Autophagy
030304 developmental biology
business.industry
ICC
Immunocytochemistry
LC3
Microtubule associated protein 1A/1B-light chain 3
PNIPAAm
Poly-N-isopropylacrylamide
medicine.disease
Cell culture
TMZ
Temozolomide
FBS
Fetal bovine serum
Cancer research
business
Zdroj: MethodsX, Vol 8, Iss, Pp 101473-(2021)
MethodsX
ISSN: 2215-0161
Popis: Rhabdomyosarcoma (RMS) is the most common pediatric soft-tissue malignant tumor. Treatment of RMS usually includes primary tumor resection along with systemic chemotherapy. Two-dimensional (2D) cell culture systems and animal models have been extensively used for investigating the potential efficacy of new RMS treatments. However, RMS cells behave differently in 2D culture than in vivo, which has recently inspired the adoption of three-dimensional (3D) culture environments. In the current paper, we will describe the detailed methodology we have developed for fabricating a 3D engineered model to study alveolar RMS (ARMS) in vitro. This model consists of a thermally cross-linked collagen disk laden with RMS cells that mimics the structural and bio-chemical aspects of the tumor extracellular matrix (ECM). This process is highly reproducible and produces a 3D engineered model that can be used to analyze the cytotoxicity and autophagy induction of drugs on ARMS cells. The most improtant bullet points are as following:•We fabricated 3D model of ARMS.•The current ARMS 3D model can be used for screening of chemotherapy drugs.•We developed methods to detect apoptosis and autophagy in ARMS 3D model to detect the mechansims of chemotherapy agents.
Graphical abstract Rhabdomyosarcoma (RMS) is the most common pediatric soft-tissue malignant tumor. Despite numerous clinical trials that have investigated the efficacy of multidrug chemotherapy regimens, outcomes for high-risk RMS patients are still poor which has inspired research into new targeted therapies that molecularly target malignant RMS cells. In order to evaluate such therapies in vitro in a realistic tumor microenvironment, we have bioengineered a cell-laden 3D tissue-engineered model with uniform geometry and an uncomplicated and reproducible biofabrication process. The transparent collagen-I hydrogel promotes cell attachment and growth and allows for analysis of the cellular response to drugs through brightfield microscopy, immunocytochemistry, and live/dead viability assays.Image, graphical abstract
Databáze: OpenAIRE
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