共培养策略在类器官研究中的应用进展

doi:10.19401/j.cnki.1007-3639.2023.03.014

中国癌症杂志 ›› 2023, Vol. 33 ›› Issue (3): 293-302.doi: 10.19401/j.cnki.1007-3639.2023.03.014

• 综述 • 上一篇

共培养策略在类器官研究中的应用进展

芦瑜¹(), 席雨梦¹, 何晓明¹, 杨少坤¹, 张佳², 王雷³, 何朝星¹(), 向柏¹()

1.河北医科大学药学院，河北石家庄 050017
2.石家庄市第四医院药剂科，河北石家庄 050035
3.河北医科大学第四医院胸外科，河北石家庄 050011

收稿日期:2022-05-13 修回日期:2022-08-05 出版日期:2023-03-30 发布日期:2023-04-17
通信作者: 何朝星（ORCID: 0000-0001-8019-0622），硕士，高级实验师；向柏（ORCID: 0000-0002-4789-641X），博士，教授，河北医科大学药学院药剂学教研室主任。
作者简介:芦瑜（ORCID: 0000-0002-0200-7973），硕士在读。
基金资助:
国家自然科学基金(81973251)

Advances in the application of co-culture strategies in organoids

LU Yu¹(), XI Yumeng¹, HE Xiaoming¹, YANG Shaokun¹, ZHANG Jia², WANG Lei³, HE Chaoxing¹(), XIANG Bai¹()

1. School of Pharmacy, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China
2. Department of Pharmacy, the Fourth Hospital of Shijiazhuang City, Shijiazhuang 050035, Hebei Province, China
3. Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei Province, China

Received:2022-05-13 Revised:2022-08-05 Published:2023-03-30 Online:2023-04-17
Contact: HE Chaoxing, XIANG Bai

摘要/Abstract

摘要：

类器官作为一种良好的体外研究模型，在生物医学领域中的应用越来越广泛。通过采用各种组织培养技术开发自组装的3D结构，类器官可重现器官固有结构中细胞的高度复杂性，因此被广泛用于研究调节机体发育和疾病的机制、高通量药物筛选及个性化治疗等。为更好地重现微环境内细胞间的相互作用，共培养策略已经扩展到更多的细胞类型，共培养策略的迅速发展使类器官技术的应用前景更加广阔，并为治疗人类疾病和再生医学开辟了新的道路。本文阐述共培养策略在类器官生成中的作用，并重点介绍不同细胞成分及微生物组与类器官共培养的应用，以期为构建开发具有更高体内模拟程度的类器官提供参考和帮助。

关键词: 共培养, 类器官, 疾病建模, 细胞间相互作用, 微环境

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

中图分类号:

R73-35+1

芦瑜, 席雨梦, 何晓明, 杨少坤, 张佳, 王雷, 何朝星, 向柏. 共培养策略在类器官研究中的应用进展[J]. 中国癌症杂志, 2023, 33(3): 293-302.

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.

图/表 3

图1

表1

免疫细胞和类器官共培养的实例"

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]

表1

表2

成纤维细胞和类器官的共培养示例"

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]

表2

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共培养策略在类器官研究中的应用进展

Advances in the application of co-culture strategies in organoids

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