Reza Bayat Mokhtari1,2, Bessi Qorri1, Manpreet Sambi1, Narges Baluch3, Sushil Kumar4, Bikul Das2,5, Myron R Szewczuk1, Herman Yeger6, Hai-Ling Margaret Cheng7,8. 1. Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON K7L 3N6, Canada. 2. Department of Experimental Therapeutics,Thoreau Laboratory for Global Health,M2D2, University of Massachusetts, Lowell, MA 01852, USA. 3. Department of Immunology and Allergy, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada. 4. QPS, Holdings LLC, Pencader Corporate Center, 110 Executive Drive, Newark, DE 19702, USA. 5. Department of Cancer and Stem Cell Biology, KaviKrishna Laboratory,GBP, Indian Institute of Technology, Guwahati 781039, India. 6. Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada. 7. The Edward S. Rogers Sr. Department of Electrical & Computer Engi-neering, Institute of Biomedical Engineering, University of Toronto, Toronto, ON M5G 1M1, Canada. 8. Translational Biology & Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto, Toronto, ON M5G 1M1, Canada.
Abstract
Therapeutic targeting of stem cells needs to be strategically developed to control tumor growth and prevent metastatic burden successfully. Breast cancer presents a unique clinical problem because of the variety of cellular subtypes present, including cancer stem cells (CSCs). The development of 3D stem-like properties of human breast tumor spheroids in stem cell factor conditioned media was investigated in orthotopic xenografts for enhanced tumorgenicity in the athymic nude rat model. MCF-7, ZR-75-1, and MDA-MB-231 breast cancer cell lines were cultured in serum-free, stem cell factor-supplemented medium under non-adherent conditions and passaged to generate 3rd generation spheroids. The spheroids were co-cultured with fetal lung fibroblast (FLF) cells before orthotopic heterotransplantation into the mammary fat pads of athymic nude rats. Excised xenografts were assessed histologically by H&E staining and immunohistochemistry for breast cancer marker (ERB1), proliferation marker (Ki67), mitotic marker (pHH3), hypoxia marker (HIF-2α), CSC markers (CD47, CD44, CD24, and CD133), and vascularization markers (CD31, CD34). Breast cancer cells cultured in stem cell factor supplemented medium generated 3D spheroids exhibited increased stem-like characteristics. The 3D stem-like spheroids co-cultured with FLF as supporting stroma reproducibly and efficiently established orthotopic breast cancer xenografts in the athymic nude rat.
Therapeutic targeting of stem cells needs to be stpan class="Species">rategically developed to control pan class="Disease">tumor growth and prevent metastatic burden successfully. Breast cancer presents a unique clinical problem because of the variety of cellular subtypes present, including cancer stem cells (CSCs). The development of 3D stem-like properties of humanbreast tumor spheroids in stem cell factor conditioned media was investigated in orthotopic xenografts for enhanced tumorgenicity in the athymic nude rat model. MCF-7, ZR-75-1, and MDA-MB-231 breast cancer cell lines were cultured in serum-free, stem cell factor-supplemented medium under non-adherent conditions and passaged to generate 3rd generation spheroids. The spheroids were co-cultured with fetal lung fibroblast (FLF) cells before orthotopic heterotransplantation into the mammary fat pads of athymic nude rats. Excised xenografts were assessed histologically by H&E staining and immunohistochemistry for breast cancer marker (ERB1), proliferation marker (Ki67), mitotic marker (pHH3), hypoxia marker (HIF-2α), CSC markers (CD47, CD44, CD24, and CD133), and vascularization markers (CD31, CD34). Breast cancer cells cultured in stem cell factor supplemented medium generated 3D spheroids exhibited increased stem-like characteristics. The 3D stem-like spheroids co-cultured with FLF as supporting stroma reproducibly and efficiently established orthotopic breast cancer xenografts in the athymic nude rat.
Entities:
Keywords:
cancer stem cells; multicellular 3D spheroid culture; orthotopic tumor xenograft
Authors: Olga Troitskaya; Diana Novak; Anna Nushtaeva; Maria Savinkova; Mikhail Varlamov; Mikhail Ermakov; Vladimir Richter; Olga Koval Journal: Int J Mol Sci Date: 2021-11-29 Impact factor: 5.923