Success rate of current human-derived gastric cancer organoids establishment and influencing factors: A systematic review and meta-analysis

Table 1 The clinical characteristic of gastric cancer organoids

Ref.	Location	Molecular subtype	Histologic classification	Differentiation	pTNM stage	Source
Yan et al[21]	Body, fundus, cardia, antrum	EBV, MSI, Mixed	Intestinal, diffuse, mixed mucinous, mixed	Moderate, poor	IA-IIIC	Surgery
Kawasaki et al[22]	NM	NM	NET, NEC	NM	NM	Surgery, biopsy
Nanki et al[23]	GEJ, corpus, antrum	MSI, CIN, GS, MSS	Intestinal, diffuse, mixed	NM	IA-IV	Surgery, biopsy, ascites puncture
Steele et al[24]	Fundus	NM	Intestinal, diffuse, signet-ring cell	Poorly differentiated	NM	Surgery
Li et al[25]	Antrum, cardia, body	NM	Intestinal, diffuse, mixed	Poor, moderate, high	IA-IV	Surgery
Zou et al[26]	NM	NM	Intestinal, mixed, SRCC	Poor, moderate	IIB-IVA	Surgery, biopsy
Li et al[27]	Antrum, corpus, cardia	NM	NM	Poor, signet-ring cell, middle	NM	Biopsy
Togasaki et al[28]	NM	GS, CIN	Diffuse	Poor, sig, muc	IIA-IV	Surgery, ascites, biopsy, autopsy
Xiao et al[29]	NM	MSI, HER2, EBV	Adenocarcinoma	Moderate to poor differentiation	III	Surgery
Seidlitz et al[30]	NM	NM	Intestinal, diffuse, mixed	NM	IIA-IV	Surgery
Gao et al[31]	NM	NM	Adenocarcinoma	NM	NM	Surgery, biopsy
Bartfeld et al[32]	Corpus	NM	NM	NM	NM	Surgery
Wang et al[33]	Cardia, corpus	NM	SCRR	Poorly differentiated	IIB-III	Biopsy
Kumar et al[34]	Antrum, distal, corpus, pylorus, cardia, proximal, peritoneum	GS, MSI, CIN	Intestinal, diffuse, mixed	NM	I-IV	Biopsy
Harada et al[35]	NM	NM	Por, tub1, tub2, muc	Differentiated, undifferentiated	IA-IIIC	Surgery
Ukai et al[36]	NM	NM	Intestinal, diffuse, tub1, tub2, muc, pap	Por	IA-IIIC	Surgery
Tong et al[37]	NM	MSS, MSI	Intestinal	NM	NM	NM
Yamaguchi et al[38]	Corpus, antrum	NM	NM	NM	NM	Surgery or ESD

pTNM: Pathological tumor-node-metastasis; NM: Not mentioned; ESD: Endoscopic submucosal dissection; NET: Neuroendocrine tumor; NEC: Neuroendocrine carcinoma; EBV: Epstein-Barr virus; CIN: Chromosomal instability; GS: Genomically stable; MSS: Microsatellite stable; MSI: Microsatellite instable; SRCC: Signet-ring cell cancer; GEJ: Gastroesophagus junction; HER2: Human epidermal growth factor receptor 2.

Table 2 The culture characteristic of included studies

Ref.	Digestion	Success/total	Morphology	Passage time	Culture medium changing time
Yan et al[21], 2018	EDTA, DL-dithiothreitol	34/68	NM	2 wk	Per 2-3 d
Kawasaki et al[22], 2020	Liberase TH, TrypLE express	3/8	NM	NM	Per 3-4 d
Nanki et al[23], 2018	Liberase TH, TrypLE express	44/59	Solid, glandular, mixed	NM	Every 3 or 4 d
Steele et al[24], 2019	Collagenase, hyaluronidase	7/10	Spherical nest, cribriform glandular	NM	Every 2 d
Li et al[25], 2022	Collagenase	12/26	Glandular, solid, cystic, grape-like, or mixed	Every 2 wk	Every 3 d
Zou et al[26], 2022	Collagenase I, II, IV	9/10	Glandular, solid, mixed	Every 6-8 d	Every 2-3 d
Li et al[27], 2023	NM	12/26	Dense morphology and no lumen. Only a few showed cystic structure and epithelial thickening	Every 1-2 wk	Every 3 d
Togasaki et al[28], 2020	Liberase TH, TrypLE express	7	NM	NM	Every 3-4 d
Xiao et al[29], 2020	Collagenase	3	Cystic	Every 2-3 d	Every 4 d
Seidlitz et al[30], 2019	EDTA, collagenase, hyaluronidase	20	Non-coherent grape-like growth pattern, compact morphology with no lumen, a single layered epithelium and cyst-like structure	Twice a week	Twice a week
Gao et al[31], 2018	Collagenase II, TrypLE	15	Gastric pit cells surrounding a central lumen	Every 5-8 d	Every 2-3 d
Bartfeld et al[32], 2015	EDTA	10	Buddings that surrounded a central lumen	Every 14 d	Every 2-3 d
Wang et al[33], 2019	NM	3	NM	NM	Every 2 d
Kumar et al[34], 2022	Collagenase	31	NM	Every 7-10 d	NM
Harada et al[35], 2021	EDTA	12	Crypt-like	Twice a week	Every 3 d
Ukai et al[36], 2020	15 mM EDTA	10	Crypt-like	Twice a week	Every 2-3 d
Tong et al[37], 2023	Not accessible	56/58	NM	NM	NM
Yamaguchi et al[38], 2022	Collagenase type 3, DNAse I	14	Cystic	NM	NM

NM: Not mentioned.

Table 3 The growth factor applied in the gastric cancer organoids culture

Ref.	Wnt	R-spondin-1	Noggin	B27	EGF	FGF-10	N-acetylcystenine	Gastrin	A83-01	FGF-2	IGF-1	Nicotinamide	Noggin	Nutlin-3	Y-27632	HGF	Other
Tong et al[37]	+	+	+	+	+	+	+	+	+	+			+
Steele et al[24]	+	+	+		+	+	+	+	+				+		+	+	SB202190, p38, MAPK inhibitor, prostaglandin E2
Nanki et al[23]	+	+	+		+	+			+				+	+
Yan et al[21]	+	+	+	+	+	+	+	+	+	+			+
Li et al[25]	+	+	+	+	+	+	+	+	+			+	+		+
Kawasaki et al[22]	+	+	+		+	+			+	+	+		+	+		+	GDNF, DAPTγ-secretase inhibitor and Palbociclib CDK4/CDK6 selective inhibitor
Togasaki et al[28]	+	+	+		+	+			+
Seidlitz et al[30]	+	+	+	+	+	+	+	+	+			+
Gao et al[31]	+	+	+		+	+		+	+			+					Hamburg, TGF, rho-associated
Bartfeld et al[32]	+	+	+		+	+		+	+			+					Hamburg, TGF, rho-associated
Wang et al[33]	+	+	+		+	+			+			+	+		+	+	Prostaglandin E2, SB202190
Kumar et al[34]	+	+	+		+	+		+	+			+	+		+	+	Prostaglandin E2, SB202190
Harada et al[35]	+	+	+	+	+	+	+		+			+	+		+		SB203580
Ukai et al[36]	+	+	+	+	+	+	+		+			+	+		+		SB203580
Yamaguchi et al[38]	+	+		+	+	+	+	+				+			+		TGF-βi

EGF: Epidermal growth factor; FGF: Fibroblast growth factor; IGF-1: Insulin-like growth factors-1; HGF: Hepatocyte growth factor; GDNF: Glialcellline-derived neurotrophic factor; TGF: Transforming growth factor.

Table 4 The quality assessment according to our adjusted Methodological Index for Non-Randomized Studies tool

Criteria	Tong et al[37], 2023	Steele et al[24], 2019	Nanki et al[23], 2018	Yan et al[21], 2018	Li et al[25], 2022	Kawasaki et al[22], 2020	Li et al[27], 2023	Zou et al[26], 2022
A clearly stated aim	2	2	2	2	2	2	2	2
Inclusion of consecutive patients	2	2	2	2	2	2	2	2
Prospective collection of data	2	2	2	2	2	2	2	2
Endpoints appropriate to the aim of the study	2	2	2	2	2	2	2	2
Follow-up period appropriate to the aim of the study	2	1	2	2	1	2	2	1
Loss to follow up less than 5%	2	1	2	2	2	2	2	1
Baseline equivalence of groups	2	2	2	2	1	1	1	2
Adequate statistical analyses	2	2	2	2	2	2	2	2
Total	16	15	16	16	14	15	15	14

The items are scored 0 (not reported), 1 (reported but inadequate) or 2 (reported and adequate). Criteria: (1) A clearly stated aim: The question addressed should be precise and relevant in the light of available literature; (2) Inclusion of consecutive patients: All patients potentially fit for diagnostic criteria of gastric cancer have been included in the study during the study period; (3) Prospective collection of data: Data were collected according to a protocol established before the beginning of the study; (4) Endpoints appropriate to the aim of the study: Unambiguous explanation of the criteria of successfully culture; (5) Follow-up period appropriate to the aim of the study: The follow-up should be sufficiently long at least for 30 d to allow the assessment of the stable construction; (6) Loss to follow up less than 5%: All gastric cancer organoids should be included in the follow up. Otherwise, the proportion fail to follow up should not exceed the proportion experiencing the major endpoint; (7) Baseline equivalence of groups: The groups should be similar regarding the criteria other than the studied endpoints. Absence of confounding factors that could bias the interpretation of the results; and (8) Adequate statistical analyses: The statistics were in accordance with the type of study.

Table 5 The gastric cancer organoids construction rate from different subgroups

Subgroup	Successful cases	Number of total	Number of study	Success rate (%)	95%CI	P value	I2
Gender
Female	14	23	4	67.0	31.1-95.7	0.31	36.07
Male	22	47	4	46.7	31.5-62.2
Tissue source
Endoscopic biopsy	16	32	3	53.7	27.1-79.5	0.28	37.35
Surgery and ESD	28	40	4	70.9	49.8-88.7
Differentiate type
Poor	27	43	3	64.6	46.0-81.3	0.106	42.54
Moderate	5	15	2	31.0	6.3-61.3
SRCC	6	16	4	32.7	0.6-76.7
pTNM
I-II	3	8	2	38.3	4.1-79.1	0.108	42.54
III-IV	18	28		65.2	45.7-82.7
Enzyme
Liberase TH, TrypLE	47	67	2	71.5	59.3-82.3	0.04	69.27
EDTA, DL-dithiothreitol	34	68	1	50.0	37.6-62.4
Collagenase	30	46	3	72.1	44.4-93.6
Growth factor
B27	103	152	3	68.7	46.9-83.3	0.94	90.53
Non-B27	55	77	3	69.5	45.2-87.8
N-acetylcysteine	111	162	4	71.9	38.2-96.2	0.95	90.52
Non-N-acetylcysteine	47	67	2	71.5	59.3-82.3
Gastrin	111	162	4	71.9	38.2-96.2	0.95	90.53
Non-gastrin	47	67	2	71.5	59.3-82.3
FGF-2	93	134	3	67.4	21.6-99.2	0.98	90.53
Non-FGF-2	65	95	3	67.9	49.2-84.2
Nutlin-3	47	67	2	71.5	59.3-82.3	0.95	90.53
Non-nutlin-3	111	162	4	71.9	38.2-96.2
Y-27632	21	36	2	58.2	41.6-75.0	0.505	90.53
Non-Y-27632	137	193	4	70.0	39.8-93.3

pTNM: Pathological tumor-node-metastasis; CI: Confidence interval; ESD: Endoscopic submucosal dissection; SRCC: Signet-ring cell cancer; FGF-2: Fibroblast growth factor-2.