Table 1.

Review of in vitro breast cancer-adipose preclinical platforms, which includes a summary of associated cell types, derivation, key findings, and the strengths and/or weaknesses of the platform demonstrated in each study.

Platform	Cell Type(s)	Human-Derived (Y/N)	Patient-Derived Adipocytes or Adipose-Derived Stromal/Stem Cells (ASCs) (Y/N)	Multi Patient Adipocytes or ASCs (Y/N)	Key Findings	Strengths and/or Weaknesses	Reference
2D Co-Culture	THP-1 Macrophages, HUVECs, and Mammary Preadipocytes	Yes	Yes	No	Macrophage vascular endothelial growth factor A (VEGFA) expression increased post co-culture. Macrophage expression of pro-angiogenic and pro-metastatic genes significantly increased post co-culture. Conditioned media derived from co-cultures promoted HUVEC endothelial tube formation	No breast cancer cells were used	[48]
	MSCs, U937, MCF-7, and MDA-MB-231	Yes	Yes	No	Macrophage paracrine activity intensifies breast cancer cell-adipocyte crosstalk	2D co-culture	[27]
	ASCs, MCF-7, T47D, and ZR75	Yes	Yes	Yes	Obesity-altered ASCs contribute to the radiation resistance observed in ER+ breast cancer	2D co-culture	[49]
	ASCs, MCF-7, and MDA-MB-231	Yes	Yes	Yes	ASCs increase breast cancer cell proliferation. ASC paracrine activity increases breast tumor cell proliferation	2D co-culture	[26]
	MDA-MB-231 and Adipocytes	No	Yes	No	MDA-MB-231 pro-inflammatory gene expression was upregulated in the presence of obese murine adipose tissue. MDA-MB-231 impacted adipocyte biosynthesis pathways	2D co-culture	[25]
	HS578Bst and ASCs	Yes	Yes	Yes	ASCs disrupted the expression of ECM maintenance-related genes and increased leptin and inflammatory marker gene expression	2D co-culture	[24]
	ASCs, MDA-MB-231, and MCF-7	Yes	Yes	Yes	ASCs do not impact breast cancer cell proliferation via direct cell contact or paracrine activity. ASCs do not significantly increase breast cancer cell EMT-related gene expression via direct cell contact	2D co-culture	[33]
3D Co-Culture	3T3-F442A, ZR75, SUM159PT, MCF-7, T47D, and MDA-MB-231	Yes-Breast Cancer Cell Lines No-Preadipocytes	No	No	Adipocytes promote the invasion of MDA-MB-231, MCF-7, and other breast cancer cell lines	No human-derived preadipocytes used	[45]
	ASCs and breast cancer cell lines	Yes	Yes	No	Demonstrates breast cancer cell-primary preadipocyte crosstalk in vitro	Sandwich white adipose tissue-breast cancer model	[59]
	Breast Adipose Tissue, ASCs, and TU-BcX-41C	Yes	Yes	Yes	Increase in cancer stem cell population when TU-BcX-41C cells are cultured in breast cancer-adipose MPS	Modified sandwich MPS model	[60]
	3T3-L1 pre-adipocytes, MDA-MB-231, MCF-7, SUM159, and HS578t	Yes	No	No	Breast cancer cell interactions with the ECM and adipocytes alter breast cancer cell MET, potentially contributing to secondary tumor formation	Used 3T3-L1 preadipocytes	[53]
	MSCs and MDA-MB-231	Yes	Yes	No	MDA-MB-231 invasion is enhanced in the presence of adipocytes and collagen matrix	Integrated a collagen plug into traditional Boyden chamber	[55]
	3T3-L1 pre-adipocytes, MDA-MB-453, MDA-MB-435S, MDA-MB-231, and MDA-MB-468	Yes	No	No	Adipocytes induce migration and invasion of breast cancer cells. Adipocytes stimulate breast cancer cells to adopt an aggressive tumor phenotype by inducing EMT-associated traits	Used 3T3-L1 preadipocytes	[28]
	Stromal vascular fraction (SVF), ASCs, and MDA-MB-231	Yes	Yes	Yes	Direct and indirect contact with adipocytes induces similar invasive behaviors in the MDA-MB-231 TNBC cell line	Enhanced cellular heterogeneity by including SVF	[29]
	Mammary adipocytes and MDA-MB-231	Yes	Yes	Yes	Breast cancer-adipocyte crosstalk is amplified by obesity. Supports the study of mammary adipocyte lipid secretion on tumor secretions and overall tumor aggressiveness in lean and obese conditions	Used a fibrin matrix in Boyden chamber	[56]
	ASCs, Fibroblasts, and MDA-MB-231	Yes	No	No	Collagen VI, a highly oncogenic collagen isoform linked to breast cancer, was decreased in the irradiated cancer co-culture. Irradiation not only makes cells ablative but also may influence the oncogenic potential of the microenvironment	Used decellularized scaffold	[57]
	Murine peri-uterine and inguinal white adipose tissue (WAT) and MDA-MB-231	Yes	No	No	Adipose tissue paracrine activity induces MET-like changes in the MDA-MB-231 TNBC cell line	Used murine WAT	[54]
Microfluidics	ASCs and MDA-MB-231	Yes	Yes	No	Statistically significant increase in MDA-MB-231 proliferation in the presence of ASCs. MDA-MB-231 cells adopt more of a mesenchymal phenotype in the presence of ASCs. Paclitaxel has reduced effectiveness in inhibiting MDA-MB-231 replication in the presence of ASCs	One ASC donor used (female, Caucasian, normal BMI)	[30]
	MCF-7 and ASCs	Yes	Yes	Yes	Predicts anastrazole sensitivity with respect to ASC BMI better than a 2D co-culture system. Primary mammary adipose stromal cells derived from obese patients exhibited increased aromatase mRNA compared to lean controls	Multiple ASC donors used	[32]
Spheroids	ASCs, MDA-MB-231, MCF-7, DT28, and HMLER3 CSC	Yes	Yes	Yes	The proportion of mammosphere-forming cells and cells expressing stem-like markers increases when in direct or indirect contact with adipocytes	Multiple ASC donors used	[62]
	Multipotent adipose-derived stem cells (MADS)-adipocytes, breast adipocytes, MCF-7, and MDA-MB-231	Yes	Yes	No	Adipocyte lipid droplet size decreases in the presence of mammospheres. UCP1 expression is dependent on adipocyte-mammosphere distance. Mammospheres produce adrenomedullin, which is critical in the interactions between adipocytes and breast cancer cells	hMADS cells isolated from young donors [63]	[64]
	ASCs, MDA_MB-231, and MCF-7	Yes	No	No	ASC C-C motif chemokine ligand 5 (CCL5) expression was elevated when co-cultured with the MDA-MB-231 TNBC cell line	One ASC donor used	[65]
	3T3-L1 pre-adipocytes, SKBR-3, MDA-MB-231, and MDA-MB-468	Yes	No	No	Adipocytes increase the invasiveness of breast cancer cells	Used 3T3-L1 preadipocytes	[43]
	ASCs, MCF10AT, MCF10A, MCF10DCIS.com, MCF10CA1a	Yes	Yes	No	ASCs promote premalignant breast cell invasions via direct cell contact. Obese ASCs have a pro-invasive effect on premalignant and malignant breast cell lines	Combination of models established in study	[66]
Bioprinting	ASCs and MDA-MB-231	Yes	No	No	MDA-MB-231 TNBC cell line induces adipose tissue ECM remodeling and lipid content modulation	Used hyaluronic acid-based bioink and extrusion-based bioprinting	[67]
	ASCs and MDA-MB-231	Yes	Yes	Yes	Adipose cells hasten the invasion and escape of tumor cells via soluble factor secretion. Tumor invasion and escape are more strongly induced by ASCs than adipocytes	Multiple demographics represented in ASC donor selection	[31]