Isolation and functional characterization of pericytes derived from hamster skeletal muscle

Ce, Mogensen; Bergner, B.; Wallner, Stefan; Ritter, Angela; Davis, Susan E.; Ninichuk, Volha; Kameritsch, Petra; Gloe, Torsten; Nagel, Wolfgang; Pohl, Ulrich

doi:10.1111/j.1748-1716.2010.02206.x

Cited by 24 publications

(21 citation statements)

References 72 publications

(149 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The rapid transmission of intercellular Ca 2+ signals from arteriolar myocytes to capillary and venular pericytes along microvessel axis, which requires depolarization, L-type Ca 2+ channels and gap junctions, has recently been shown in ureteric microvascular networks in situ [15]. The intercellular Ca 2+ signals between the pericytes of isolated vasa recta renis have also been recently observed [62]. However, it is not clear whether an axial spread of intercellular Ca 2+ signals from myocytes to pericytes involves the endothelium.…”

Section: Vgcc-dependent Intercellular Ca2+ Signallingmentioning

confidence: 98%

Calcium Signalling in Pericytes

Burdyga

Borysova²

2014

J Vasc Res

View full text Add to dashboard Cite

Recent advances in pericyte research have contributed to our understanding of the physiology and pathophysiology of microvessels. The microvasculature consists of arteriolar and venular networks located upstream and downstream of the capillaries. Arterioles are surrounded by a monolayer of spindle-shaped myocytes, while terminal branches of precapillary arterioles, capillaries and all sections of postcapillary venules are encircled by a monolayer of morphologically diverse pericytes. There are physiological differences in the response of pericytes and myocytes to vasoactive molecules, suggesting that these two vascular cell types could have different functional roles in the regulation of local blood flow. The contractile activity of pericytes and myocytes is controlled by changes of cytosolic free Ca²⁺ concentration. In this short review, we summarize our results and those of other authors on the contractility of pericytes and their Ca²⁺ signalling. We describe results regarding sources of Ca²⁺ and mechanisms of Ca²⁺ release and Ca²⁺ entry in control of the spatiotemporal characteristics of the Ca²⁺ signals in pericytes.

show abstract

Section: Vgcc-dependent Intercellular Ca2+ Signallingmentioning

confidence: 98%

Calcium Signalling in Pericytes

Burdyga

Borysova²

2014

J Vasc Res

View full text Add to dashboard Cite

show abstract

“…To date, interest in this aspect has concentrated primarily on intercellular communication within endothelial tissue (28,31,76,78), and there is already good evidence for electrophysiological mechanisms in this respect (11,26,28,58,88,92).…”

Section: Functional Characterization Of Pericytesmentioning

confidence: 99%

“…While considerable attempts have also been made to culture pericytes (18,22,35,43,52,58,60,62,74,84,95), these have not found comparably broad acceptance. These cells must be isolated from microvascular preparations, and the latter must first be separated proteolytically from their surrounding parenchyma, connective tissue, and lymphatic system, naturally always complex.…”

mentioning

confidence: 97%

Isolation, bulk cultivation, and characterization of coronary microvascular pericytes: the second most frequent myocardial cell type in vitro

Nees

Weiss

Senftl

et al. 2012

American Journal of Physiology-Heart and Circulatory Physiology

View full text Add to dashboard Cite

Densely arranged pericytes engird the endothelial tube of all coronary microvessels. Since the experimental access to these abundant cells in situ is difficult, a prerequisite for broader investigation is the availability of sufficient numbers of fully differentiated pericytes in homogenous culture. To reach this goal, we applied strictly standardized cell isolation techniques, optimized culture methods and specific histological staining. Approximately 1,000-fold enriched pericytes were proteolytically detached from highly purified coronary microvascular networks (density gradient centrifugation) of eight mammalian species including human. Addition of species-autologous fetal or neonatal serum (10-20% vol/vol) was a precondition for longer term survival of homogenous pericyte cultures. This ensured optimal growth (doubling time <14 h) and full expression of pericyte-specific markers. In 3-mo, 10(10) pericytes (15 g) could be cultivated from 1 bovine heart. Pericytes could be stored in liquid N(2), recultured, and passaged repeatedly without loss of typical features. In cocultures with EC or vascular smooth muscle cells, pericytes transferred fluorescent calcein to each other and to EC via their antler-like extensions, organized angiogenetic sprouting of vessels, and rapidly activated coagulation factors X and II via tissue factor and prothrombinase. The interconnected pericytes of the coronary system are functionally closely correlated with the vascular endothelium and may play key roles in the adjustment of local blood flow, the regulation of angiogenic processes, and the induction of procoagulatory processes. Their successful bulk cultivation enables direct experimental access under defined in vitro conditions and the isolation of pericyte specific antigens for the production of specific antibodies.

show abstract

“…Most established methods for isolating blood/tissue barrier cells, including ECs and PCs, were developed on large tissue masses, such as brain, retina, skeletal muscle, skin, and fetal tissues (Abbott et al, 2012; Bryan et al, 2008; Katyshev et al, 2012; Maier et al, 2010; Mogensen et al, 2011; Nees et al, 2012; Siow, 2012). These methods are typically complex procedures, involving multiple steps of enzymatic digestion and gradient density centrifugation.…”

Section: Discussionmentioning

confidence: 99%

“…However, these research tools have been of limited use in studying the BLB due to the difficulty of isolating BLB component cells from the vestibular system. Although different methods of isolation and culture of barrier cells from the brain, retina, skeletal muscle, skin, and fetal tissues have been successfully used to obtain barrier component cells (Bryan et al, 2008; Crisan et al, 2008; Mogensen et al, 2011; Sundberg et al, 2002), most of the methods are time-consuming and involve multiple steps of enzymatic digestion, gradient density centrifuging, and glass bead or magnetic separation (Bowman et al, 1983; Bowman et al, 1981; Ohtsuki et al, 2007; Stins et al, 1997). The techniques are usually performed in non-cochlear tissues from rat (Ohtsuki et al, 2007), porcine (Mischeck et al, 1989), or bovine models (Ryan, 1984) where tissue volume is not limited(Ballarin et al, 2012; Leppens et al, 1996; Xie et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system

et al. 2017

View full text Add to dashboard Cite

The vestibular blood-labyrinth barrier (BLB) is comprised of perivascular-resident macrophage-like melanocytes (PVM/Ms) and pericytes (PCs), in addition to endothelial cells (ECs) and basement membrane (BM), and bears strong resemblance to the cochlear BLB in the stria vascularis. Over the past few decades, in vitro cell-based models have been widely used in blood-brain barrier (BBB) and blood-retina barrier (BRB) research, and have proved to be powerful tools for studying cell-cell interactions in their respective organs. Study of both the vestibular and strial BLB has been limited by the unavailability of primary culture cells from these barriers. To better understand how barrier component cells interact in the vestibular system to control BLB function, we developed a novel culture medium–based method for obtaining EC, PC, and PVM/M primary cells from tiny explants of the semicircular canal, sacculus, utriculus, and ampullae tissue of young mouse ears at post-natal age 8 - 12 d. Each phenotype is grown in a specific culture medium which selectively supports the phenotype in a mixed population of vestibular cell types. The unwanted phenotypes do not survive passaging. The protocol does not require additional equipment or special enzyme treatment. The harvesting process takes less than 2 h. Primary cell types are generated within 7 - 10 d. The primary culture ECs, PCs, and PVM/M shave consistent phenotypes more than 90% pure after two passages (~ 3 weeks). The highly purified primary cell lines can be used for studying cell-cell interactions, barrier permeability, and angiogenesis.

show abstract

Isolation and functional characterization of pericytes derived from hamster skeletal muscle

Abstract: We established a culture method for PC from skeletal muscle. A first functional characterization revealed properties which potentially enable these cells to generate hyperpolarizing signals and to communicate them to endothelial cells.

Cited by 24 publications

References 72 publications

Calcium Signalling in Pericytes

Calcium Signalling in Pericytes

Isolation, bulk cultivation, and characterization of coronary microvascular pericytes: the second most frequent myocardial cell type in vitro

Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system

Contact Info

Product

Resources

About