Department of Immunology, Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, R3E 0T5, Canada.
Research Institute of Oncology & Hematology, CancerCare Manitoba, Winnipeg, MB, R3E 0V9, Canada.
Stem Cell Res Ther. 2018 Oct 4;9(1):264. doi: 10.1186/s13287-018-0994-y.
Normal human breast epithelial cells are maintained by the proliferation and differentiation of different human breast epithelial progenitors (HBEPs). However, these progenitor subsets can only be obtained at low frequencies, limiting their further characterization. Recently, it was reported that HBEPs can be minimally expanded in Matrigel cocultures with stromal feeder cells. However, variability of generating healthy feeder cells significantly impacts the effective expansion of HBEPs.
Here, we report a robust feeder cell-free culture system for large-scale expansion of HBEPs in two-dimensional cultures.
Using this cell culture system HBEPs can be exponentially expanded as bulk cultures. Moreover, purified HBEP subtypes can also be separately expanded using our cell culture system. The expanded HBEPs retain their undifferentiated phenotype and form distinct epithelial colonies in colony forming cell assays.
The availability of a culture system enabling the large-scale expansion of HBEPs facilitates their application to screening platforms and other cell-based assays.
正常的人类乳腺上皮细胞由不同的人类乳腺上皮祖细胞（HBEP）的增殖和分化维持。然而，这些祖细胞亚群只能以低频率获得，限制了它们的进一步表征。最近，有报道称 HBEP 可以在与基质饲养细胞的 Matrigel 共培养中最小限度地扩增。然而，生成健康饲养细胞的可变性显著影响 HBEP 的有效扩增。
在这里，我们报告了一种用于二维培养中大规模扩增 HBEP 的无饲养细胞的稳健培养系统。
使用这种细胞培养系统，HBEP 可以作为批量培养进行指数级扩增。此外，使用我们的细胞培养系统还可以分别扩增纯化的 HBEP 亚型。扩增的 HBEP 保留其未分化的表型，并在集落形成细胞测定中形成独特的上皮集落。
可大规模扩增 HBEP 的培养系统的可用性促进了它们在筛选平台和其他基于细胞的测定中的应用。
