Breast Cancer Stem Cell Culture and Enrichment Using Poly(ε-Caprolactone) Scaffolds

Abstract

Abstract:The cancer stem cell (CSC) population displays self-renewal capabilities, resistance to conventional therapies, and a tendency to post-treatment recurrence. Increasing knowledge about CSCs' phenotype and functions is needed to investigate new therapeutic strategies against the CSC population. Here, poly(ε-caprolactone) (PCL), a biocompatible polymer free of toxic dye, has been used to fabricate scaffolds, solid structures suitable for 3D cancer cell culture. It has been reported that scaffold cell culture enhances the CSCs population. A RepRap BCN3D+ printer and 3 mm PCL wire were used to fabricate circular scaffolds. PCL design and fabrication parameters were first determined and then optimized considering several measurable variables of the resulting scaffolds. MCF7 breast carcinoma cell line was used to assess scaffolds adequacy for 3D cell culture. To evaluate CSC enrichment, the Mammosphere Forming Index (MFI) was performed in 2D and 3D MCF7 cultures. Results showed that the 60˝scaffolds were more suitable for 3D culture than the 45˝and 90˝ones. Moreover, 3D culture experiments, in adherent and non-adherent conditions, showed a significant increase in MFI compared to 2D cultures (control). Thus, 3D cell culture with PCL scaffolds could be useful to improve cancer cell culture and enrich the CSCs population.

A comparison study between electrospun polycaprolactone and piezoelectric poly(3-hydroxybutyrate-co-3-hydroxyvalerate) scaffolds for bone tissue engineering

“…Wang M, Cheng X, Zhu W, Holmes B, Keidar M, Zhang LG Tissue Eng Part A. 2014 Mar; 20(5-6):1060-71; G. Kumar, et al Freeform Fabricated Scaffolds with Roughened Struts that Enhance both Stem Cell Proliferation and Differentiation by Controlling Cell Shape, Biomaterials, 2012, 33 (16): 4022 -4030], it was shown that high surface roughness of scaffolds and wettability enhance osteogenic differentiation of hMSCs on scaffolds. With respect to reviewer comments, we described this issue more clearly and added more references, confirming that statement.…”

“…The viability of hMSCs on scaffolds at 1 day of culturing was determined using a cell detachment protocol, described in [16]. The culture medium was removed with a micropipette.…”

Chemoresistance of Lung Cancer Cells: 2D and 3D In Vitro Models for Anticancer Drug Screening

“…Over the course of time, researchers have tried to improve upon the existing protocol of sphere formation in order to make it more efficient, consistent, reliable and physiologically relevant. Various types of scaffolds consisting of chitosan-alginate, collagen, alginate and agarose have been used as 3D matrices to substitute low attachment plates [69][70][71][72]. In 2014, Cao et al generated tumorspheres from primary neuroblastoma cells driven from MYCN transgenic mice using a medium supplemented with fetal bovine serum (FBS) and β-mercaptoethanol.…”

Influence of the Fabrication Accuracy of Hot-Embossed PCL Scaffolds on Cell Growths

“…In Causa et al (2006) authors described how a PCL scaffold, reinforced with hydroxyapatite (HA) particles, is a good solution for the regeneration of bones promoting the proliferation of human osteoblasts and for the regeneration of cartilage tissue (Wei et al, 2015;Zhang et al, 2016). Regarding new effective 3D cell culture applications, Palomeras et al (2016) used the PCL scaffolds for the in vitro growth of cancer stem cells (CSCs). These cell type represents a minor subpopulation in solid tumors able to drive tumor progression and metastatization.…”

“…These cell type represents a minor subpopulation in solid tumors able to drive tumor progression and metastatization. CSCs are difficult to culture in 2D systems and Palomeras et al (2016) successfully demonstrated their enrichment due to seeding them on a PCL scaffold. Their 3D PCL scaffolds resulted a useful way to enrich and propagate CSCs for further investigation targeted to this malignant subpopulation.…”