title: Differentiation dynamics of mammary epithelial stem cells from Korean holstein dairy cattle under ECM-free conditions.
abstract: The "stem cells" are commonly defined as "cells capable of self-renewal through replication and differentiating into specific lineages". The mammary gland contains functional stem/progenitor cells. The current study was planned with the objectives to study the differentiation dynamics of Korean Holstein mammary epithelial stem cells (KHMESCs) under the optimum culture conditions. Lineage negative KHMESCs isolated from mammary tissue of lactating cows have shown the typical differentiation dynamics with formation of lobulo-alveolar structures in in vitro culture. This suggests the existence of bipotential mammary epithelial stem cells in the mammary gland. The strong mRNA expression of pluripotency factors indicates stemness, whereas expression of milk protein genes and epithelial cell-specific gene indicate their differentiation capabilities. Further, immunostaining results have shown the differentiation capabilities of KHMESCs into both luminal and basal lineages under the extracellular matrix (ECM, matrigel) free environment. However, under matrigel, the differentiation process was comparatively higher than without matrigel. Immunostaining results also suggested that differentiated cells could secrete milk proteins such as β-casein. To our knowledge, these data represent the first report on the differentiation dynamics and establishment of mammary epithelial stem cells from Korean Holstein with typical stemness properties. It was observed that isolated KHMESCs had normal morphology, growth pattern, differentiation ability, cytogenetic and secretory activity even without ECM. Therefore, it is concluded that established KHMESCs could be used for further studies on Korean Holstein dairy cows related to lactation studies, as non-GMO animal bioreactors and stem cell-based management of bovine mastitis including post-mastitis damage.
authors.affiliation: a Faculty of Biotechnology, Department of Animal Biotechnology , Jeju National University , Jeju , Republic of Korea.
authors.forename: Neelesh
authors.initials: N
authors.lastname: Sharma
authors.affiliation: a Faculty of Biotechnology, Department of Animal Biotechnology , Jeju National University , Jeju , Republic of Korea.
authors.forename: Jeong Hyun
authors.initials: JH
authors.lastname: Kim
authors.affiliation: a Faculty of Biotechnology, Department of Animal Biotechnology , Jeju National University , Jeju , Republic of Korea.
authors.forename: Simrinder Singh
authors.initials: SS
authors.lastname: Sodhi
authors.affiliation: a Faculty of Biotechnology, Department of Animal Biotechnology , Jeju National University , Jeju , Republic of Korea.
authors.forename: Do Huynh
authors.initials: do H
authors.lastname: Luong
authors.affiliation: b Animal Genetic Resources Station , National Institute of Animal Science, Rural Development Administration , Namwon , Republic of Korea.
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Kim
authors.affiliation: a Faculty of Biotechnology, Department of Animal Biotechnology , Jeju National University , Jeju , Republic of Korea.
authors.forename: Sung Jong
authors.initials: SJ
authors.lastname: Oh
authors.affiliation: a Faculty of Biotechnology, Department of Animal Biotechnology , Jeju National University , Jeju , Republic of Korea.
authors.forename: Dong Kee
authors.initials: DK
authors.lastname: Jeong
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D019070:Cell Lineage / Q000235:genetics / Q000502:physiology; D049109:Cell Proliferation / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen; D004338:Drug Combinations; D004847:Epithelial Cells / Q000166:cytology*; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D007700:Kinetics; D007774:Lactation; D007797:Laminin; D008321:Mammary Glands, Animal / Q000166:cytology*; D046529:Microscopy, Electron, Transmission; D008894:Milk Proteins / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D011509:Proteoglycans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D004338:Drug Combinations; D007797:Laminin; D008894:Milk Proteins; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen
keywords: KHMESCs; differentiation dynamics; luminal cells; matrigel; myoepithelial cells