OBJECTIVE: Pericytes are critical cellular components of the microvasculature that play a major role in vascular development and pathologies, yet their study has been hindered by lack of a standardized method for their isolation and growth. Here we report a method for culturing human pericytes from a readily available tissue source, placenta, and provide a thorough characterization of resultant cell populations. METHODS: We developed an optimized protocol for obtaining pericytes by outgrowth from microvessel fragments recovered after enzymatic digestion of human placental tissue. We characterized outgrowth populations by immunostaining, by gene expression analysis, and by functional evaluation of cells implanted in vivo. RESULTS: Our approach yields human pericytes that may be serially expanded in culture and that uniformly express the cellular markers NG2, CD90, CD146, alpha-SMA, and PDGFR-beta, but lack markers of smooth muscle cells, endothelial cells, and leukocytes. When co-implanted with human endothelial cells into C.B-17 SCID/bg mice, human pericytes invest and stabilize developing human endothelial cell-lined microvessels. CONCLUSIONS: We conclude that our method for culturing pericytes from human placenta results in the expansion of functional pericytes that may be used to study a variety of questions related to vascular biology.
OBJECTIVE: Pericytes are critical cellular components of the microvasculature that play a major role in vascular development and pathologies, yet their study has been hindered by lack of a standardized method for their isolation and growth. Here we report a method for culturing human pericytes from a readily available tissue source, placenta, and provide a thorough characterization of resultant cell populations. METHODS: We developed an optimized protocol for obtaining pericytes by outgrowth from microvessel fragments recovered after enzymatic digestion of human placental tissue. We characterized outgrowth populations by immunostaining, by gene expression analysis, and by functional evaluation of cells implanted in vivo. RESULTS: Our approach yields human pericytes that may be serially expanded in culture and that uniformly express the cellular markers NG2, CD90, CD146, alpha-SMA, and PDGFR-beta, but lack markers of smooth muscle cells, endothelial cells, and leukocytes. When co-implanted with human endothelial cells into C.B-17 SCID/bg mice, human pericytes invest and stabilize developing human endothelial cell-lined microvessels. CONCLUSIONS: We conclude that our method for culturing pericytes from human placenta results in the expansion of functional pericytes that may be used to study a variety of questions related to vascular biology.
Authors: Natalia Kosyakova; Derek D Kao; Maria Figetakis; Francesc López-Giráldez; Susann Spindler; Morven Graham; Kevin J James; Jee Won Shin; Xinran Liu; Gregory T Tietjen; Jordan S Pober; William G Chang Journal: NPJ Regen Med Date: 2020-01-06
Authors: Alexander Birbrair; Tan Zhang; Zhong-Min Wang; Maria Laura Messi; Grigori N Enikolopov; Akiva Mintz; Osvaldo Delbono Journal: Stem Cells Dev Date: 2013-04-27 Impact factor: 3.272
Authors: William G Chang; Jillian W Andrejecsk; Martin S Kluger; W Mark Saltzman; Jordan S Pober Journal: Cardiovasc Res Date: 2013-09-16 Impact factor: 10.787
Authors: Lingling Neng; Wenjing Zhang; Ahmed Hassan; Marcin Zemla; Allan Kachelmeier; Anders Fridberger; Manfred Auer; Xiaorui Shi Journal: Nat Protoc Date: 2013-03-14 Impact factor: 13.491