Advances in the application of co-culture strategies in organoids

doi:10.19401/j.cnki.1007-3639.2023.03.014

Abstract

Abstract:

As a good in vitro research model, organoids are more and more widely used in the biomedical field. By developing self-assembled 3D structures using various tissue culture techniques, organoids can rebuild the high complexity of cells in the inherent structure of the organ, and are therefore unanimously used to study mechanisms regulating body development and disease, high-throughput drug screening, and personalized treatment and so on. To better recapitulate cell-to-cell interactions within the microenvironment, co-culture strategies have been extended to more cell types, and their rapid development offers broader prospects for organoids and paves the way for the treatment of human diseases and regenerative medicine. This review discussed the role of co-culture strategies in organoid generation, and focused on the application of various cellular components and microorganisms in organoid construction, thereby providing reference and help for scholars to construct and develop organoids with a higher degree of in vivo simulation.

Key words: Co-culture, Organoid, Disease modeling, Cell-to-cell interactions, Microenvironment

CLC Number:

R73-35+1

LU Yu, XI Yumeng, HE Xiaoming, YANG Shaokun, ZHANG Jia, WANG Lei, HE Chaoxing, XIANG Bai. Advances in the application of co-culture strategies in organoids[J]. China Oncology, 2023, 33(3): 293-302.

Figures/Tables 3

Fig. 1

Tab. 1

Example of co-culture of immune cells and organoids"

Organoid type	Components used	Platform and matrix	Type of disease	Main outcomes	Treatment method	Reference
Human-derived autologous organoid	DC and the patient’s CD8⁺ T cells	Cell-basement membrane matrix	Gastric cancer	Predicting PD-L1 treatment efficacy and patient prognosis in cancer	Immune checkpoint	[8]
Murine and human PDAC autologous organoid	CTL/MDSC	Matrigel	PC	The efficacy of combination therapy and targeted therapy was tested	PD-1 inhibition and MDSC depletion	[9-10]
Primary organoids derived from surgical specimens	T lymphocytes	Matrigel	CRC	CEA-TCB’s performance against three-dimensional tumor organoids	CEA-CD3 T cell bispecific antibody	[11]
Patient-derived organoid	T cell	Matrigel	Melanoma	Testing the migration and cytotoxic effector function of expanded γδT cells in patient- derived melanoma organoid	Adoptive immunotherapy	[12]
An early-stage human brain organoid	Embryonal CNS tumors and Zika virus	Matrigel	CNS	Analysis of the oncolytic role of Zika virus as an oncolytic virus	Oncolytic virus	[13]
Autologous tumor organoid	Human PBMC-derived DC	BME 2	BC	The tumor suppressor and immune-activating effects of HELA-Exos were explored in patient-derived organoids	Cancer vaccine	[14]
Human epithelial colon organoid lines derived from rectal biopsy samples	Cryopreserved primary human CD4⁺ T cells	Transwell	IBD	Determining the mechanism by which IL-22 regulates intestinal epithelial cell function	—	[16]
The epithelial layer from fetal ISC	Fetal CD4⁺ Tem cells	Matrigel	NEC	TNF-α-producing CD4⁺ T cells promote mucosal development in the fetal gut but also mediate inflammation during preterm birth	—	[17]
Murine small intestine organoids	ILC1	Matrigel	Inflammation	ILC1 may exacerbate fibrosis and tumor growth when enriched in inflamed patient tissue	—	[19]
PDO	PBMC	Matrigel	GC	Dexamethasone can increase the sensitivity of ICI in coculture system	Immune checkpoint	[21]
Autologous tumor organoids	Autologous peripheral blood lymphocytes	Geltrex	CRC or NSCLC	This enables the establishment of ex vivo test systems for T-cell-based immunotherapy at the level of the individual patient	—	[22]
Tissue-derived human gastric organoids	MoDC	Microfluidics and V-ORG-3	—	Studying DC-epithelial cell interactions in the human stomach	—	[24]
Murine intestinal tumor organoids	IEL	Matrigel	—	Abundant IEL in the small intestine may help reduce the number of tumors	—	[25]

Tab. 1

Tab. 2

Example of co-culture of fibroblasts and organoids"

Organoid type	Components used	Platform and matrix	Main outcomes	Detection technology	Reference
CRC PDO	CRC-derived CAF	Matrigel	Some oncogenic signal cascades induced by cell-cell interaction between organoids and CAF were mediated by the signal pathways related to immune responses	DNA microarray; NCC oncopanel	[30]
CRC PDO	CAF	Hyaluronan-gelatin hydrogel	Co-culture of CRC PDO with CAF resulted in enhanced PDO growth and drug resistance	RNA- and whole-exome sequencing; Immunostaining and immunohistochemistry; Viability staining	[31]
Primary liver tumor-derived organoids	CAF	Transwell	CAF promoted tumor organoid growth via paracrine signaling. Vice versa, cancer cells secrete paracrine factors regulating CAF physiology	Quantitative real-time reverse-transcription PCR; Immunohistochemistry and immunofluorescence	[32]
OSCC PDO	Primary CAF	Matrigel	Notch activity contributes to biogenesis of OSCC-associated fibroblasts	Bright phase and fluorescence imaging; Immunohistochemical analysis	[34]
Human iPSC-derived HLOs (LUAD)	Human fetal fibroblast	Matrigel	HLOs that overexpressed HER2 transformed to tumor-like structures similar to atypical adenomatous hyperplasia	RNA sequence analysis; Immunocytochemistry; Immunohistochemistry; EdU assay; Real-time-quantitative reverse-transcription PCR	[35]
Generate organoids from PC-9 cells	Podoplanin (+) CAF	StemPro human embryonic stem cell serum-free medium	Growth-promoting effect of podoplanin (+) CAF in cancer cells	Immunofluorescence; Immunohistochemical	[43]
CRC PDO	CRC-derived CAF	GelMA	Acoustic 3D bioprinting technology can be widely used for establishing various microtissues for modeling cancer invasion and other diseases	Immunofluorescence staining; Staining of tissue sections	[36]
PDAC PDO	Human CAF	Matrigel	CAF maintain the differentiated PDAC phenotype through secreting niche factors and induce distinct drug responses	Immunohistochemistry and immunofluorescent staining; Transcriptome analysis; RNA extraction and quantitative reverse-transcription PCR	[37]
Organoid models of breast cancer cell lines	CD36⁺ fibroblasts	Matrigel	Fibroblasts inhibit organoid growth and normalizes basal and lateral polarities	Brightfield or immunofluorescent microscopy and robust image analysis	[38]
PDAC tumor organoids	CAF	Transwell; Matrigel	CAF confer gemcitabine resistance of PDAC cells induced by CAF-derived hepatocyte growth factor	Immunofluorescence staining	[40]
Primary CRC organoids derived from surgical specimens	T cells and autologous CAF	Matrigel	Testing the costimulatory effect of a fibroblast activating protein (FAP)-targeted 4-1BBL bispecific antibody fusion protein currently in clinical trials	Confocal videomicroscopy	[11]
LNCaP or PC3 organoids	Prostate cancer-derived CAF	Matrigel	Targeting AR/FlnA complexes by stitching peptides provides a potential new strategy for PC therapy	PCR; IP; Co-IP; Rac pull-down assay and Western blot	[42]

Tab. 2

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