Purification and Culture of Human Blood Vessel–Associated Progenitor Cells

Abstract

Multilineage progenitor cells, diversely designated as MSC, MAPC, or MDSC, have been previously extracted from long‐term cultures of fetal and adult organs (e.g., bone marrow, brain, lung, pancreas, muscle, adipose tissue, and several others). The identity and location, within native tissues, of these elusive stem cells are described here. Subsets of endothelial cells and pericytes, which participate in the architecture of human blood vessels, exhibit, following purification to homogeneity, developmental multipotency. The selection from human tissues, by flow cytometry using combinations of positive and negative cell surface markers, of endothelial and perivascular cells is described here. In addition, a rare subset of myoendothelial cells that express markers of both endothelial and myogenic cell lineages and exhibit dramatic myogenic and cardiomyogenic potential has been identified and purified from skeletal muscle. The culture conditions amenable to the long‐term proliferation of these blood vessel–associated stem cells in vitro are also described. Curr. Protoc. Stem Cell Biol. 4:2B.2.1‐2B.2.13. © 2008 by John Wiley & Sons, Inc.

Functional muscle regeneration with combined delivery of angiogenesis and myogenesis factors

“…S2 E and F). However, the enhanced myogenic regeneration in response to VEGF and VEGF/IGF sustained delivery could also be explained by recent findings (20)(21)(22) suggesting the existence of myogenic precursors distinct from satellite cells (e.g., pericytederived cells and myoendothelial cells) endowed with multilineage potential, including high muscle regenerative potential. Stimulation of angiogenesis may increase the pool of myogenic stem cells that are available to drive muscle regeneration.…”

“…Several recent studies have demonstrated that VEGF leads to better maintenance of skeletal muscle tissue in mouse models of muscular dystrophy, suggesting that VEGF may also play a role in muscle regeneration after injury (17)(18)(19). Other findings highlight the possibility that VEGF's ability to promote vascularization could increase the availability of blood vessel-associated stem cells capable of participating in muscle regeneration (20)(21)(22). However, a direct improvement in muscle regeneration has not been documented.…”

Neural differentiation of rat aorta pericyte cells

“…The determination of the origin and identity of SCs together with their niches in adult tissue also provides important information on their participation in endogenous tissue regeneration and their possible applications as pluri-or multipotent cells in cellular regeneration therapy (6,7,(27)(28)(29)(30)(31)(32)(33). Evidence accumulated in the last few years show that the adult macro-and microvessels contain multi-or pluripotent SCs, including MSCs, and/or pericytes, as well as hemopoietic SCs, and lineage committed progenitors, such as vascular walls endothelial progenitor and Sca-1 1 smooth muscle progenitor cells (6,7,23,(29)(30)(31)(33)(34)(35)(36). Pericyte SCs have been isolated from different vascular tissues including abdominal adipose, bone marrow, dental pulp, and umbilical cord (6,7,23,(29)(30)(31)(33)(34)(35).…”

“…Evidence accumulated in the last few years show that the adult macro-and microvessels contain multi-or pluripotent SCs, including MSCs, and/or pericytes, as well as hemopoietic SCs, and lineage committed progenitors, such as vascular walls endothelial progenitor and Sca-1 1 smooth muscle progenitor cells (6,7,23,(29)(30)(31)(33)(34)(35)(36). Pericyte SCs have been isolated from different vascular tissues including abdominal adipose, bone marrow, dental pulp, and umbilical cord (6,7,23,(29)(30)(31)(33)(34)(35). Actual discussions of the role of pericytes as SCs propose that pericytes are a source for MSC (29).…”

Comparação do potencial terapêutico de células mesenquimais e pericitos em modelo murino de distrofia muscular

“…Apesar disso, observou-se um efeito benéfico nos parâmetros fisiológicos dos animais tratados, sugerindo que as células estariam participando da manutenção ou regeneração muscular através da liberação de fatores solúveis, o que é chamado de efeito parácrino (Gharaibeh et al 2011;English et al 2010;Singer & Caplan 2011;Kean et al 2013), e sugere que existam outros mecanismos in vivo de ação das células-tronco mesenquimais, como propriedades antiinflamatórias, anti-fibróticas e imunomodulatórias (Ichim et al 2010;Jorgensen 2010;Gharaibeh et al 2011;Pinheiro et al 2012;Griffin et al 2013). Outra hipótese sugere que fatores liberados pelas células injetadas estimulariam a angiogênese, o que aumentaria a disponibilidade de progenitores associados aos vasos, já que acredita-se que as CTA são derivadas destes (Crisan, Huard, et al 2008;Crisan, Yap, et al 2008;Dellavalle et al 2011;Murphy et al 2013 (Crisan, Yap, et al 2008;Schugar et al 2009;Zimmerlin et al 2010;Park et al 2011). Em seguida, outro grupo conseguiu observar a participação dessas células na regeneração muscular in vivo (Dellavalle et al 2007), bem como o papel natural dessas células na formação do tecido muscular ao longo do desenvolvimento (Dellavalle et al 2011).…”