Selective isolation and characterization of primary cells from normal breast and tumors reveal plasticity of adipose derived stem cells

Weigand A, Boos A, Tasbihi K, Beier J, Dalton PD, Schrauder M, Horch RE, Beckmann M, Strissel P, Strick R (2016)

Publication Type: Journal article

Publication year: 2016

Journal

Book Volume: 18

Pages Range: 32

Journal Issue: 1

DOI: 10.1186/s13058-016-0688-2

Abstract

There is a need to establish more cell lines from breast tumors in contrast to immortalized cell lines from metastatic effusions in order to represent the primary tumor and not principally metastatic biology of breast cancer. This investigation describes the simultaneous isolation, characterization, growth and function of primary mammary epithelial cells (MEC), mesenchymal cells (MES) and adipose derived stem cells (ADSC) from four normal breasts, one inflammatory and one triple-negative ductal breast tumors.A total of 17 cell lines were established and gene expression was analyzed for MEC and MES (n = 42) and ADSC (n = 48) and MUC1, pan-KRT, CD90 and GATA-3 by immunofluorescence. DNA fingerprinting to track cell line identity was performed between original primary tissues and isolates. Functional studies included ADSC differentiation, tumor MES and MEC invasion co-cultured with ADSC-conditioned media (CM) and MES adhesion and growth on 3D-printed scaffolds.Comparative analysis showed higher gene expression of EPCAM, CD49f, CDH1 and KRTs for normal MEC lines; MES lines e.g. Vimentin, CD10, ACTA2 and MMP9; and ADSC lines e.g. CD105, CD90, CDH2 and CDH11. Compared to the mean of all four normal breast cell lines, both breast tumor cell lines demonstrated significantly lower ADSC marker gene expression, but higher expression of mesenchymal and invasion gene markers like SNAI1 and MMP2. When compared with four normal ADSC differentiated lineages, both tumor ADSC showed impaired osteogenic and chondrogenic but enhanced adipogenic differentiation and endothelial-like structures, possibly due to high PDGFRB and CD34. Addressing a functional role for overproduction of adipocytes, we initiated 3D-invasion studies including different cell types from the same patient. CM from ADSC differentiating into adipocytes induced tumor MEC 3D-invasion via EMT and amoeboid phenotypes. Normal MES breast cells adhered and proliferated on 3D-printed scaffolds containing 20 fibers, but not on 2.5D-printed scaffolds with single fiber layers, important for tissue engineering.Expression analyses confirmed successful simultaneous cell isolations of three different phenotypes from normal and tumor primary breast tissues. Our cell culture studies support that breast-tumor environment differentially regulates tumor ADSC plasticity as well as cell invasion and demonstrates applications for regenerative medicine.

Authors with CRIS profile

Involved external institutions

Julius-Maximilians-Universität Würzburg Germany (DE)

How to cite

APA:

Weigand, A., Boos, A., Tasbihi, K., Beier, J., Dalton, P.D., Schrauder, M.,... Strick, R. (2016). Selective isolation and characterization of primary cells from normal breast and tumors reveal plasticity of adipose derived stem cells. Breast Cancer Research, 18(1), 32. https://dx.doi.org/10.1186/s13058-016-0688-2

MLA:

Weigand, Annika, et al. "Selective isolation and characterization of primary cells from normal breast and tumors reveal plasticity of adipose derived stem cells." Breast Cancer Research 18.1 (2016): 32.

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