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

Jinhui Zhang, Songlin Chen, Jing Cai, Zhiqiang Hou, Xiaohan Wang, Allan Kachelmeier, Xiao Shi

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
JournalHearing Research
DOIs
StateAccepted/In press - Jun 27 2016

Fingerprint

Pericytes
Melanocytes
Culture Media
Endothelial Cells
Macrophages
Inner Ear
Phenotype
Cell Communication
Labyrinth Vestibule
Stria Vascularis
Semicircular Canals
Primary Cell Culture
Cochlea
Cellular Structures
Blood-Brain Barrier
Basement Membrane
Ear
Retina
Permeability
Cell Membrane

Keywords

  • Blood-labyrinth barrier
  • Endothelial cell
  • Mouse cochlear vestibular system
  • Pericyte
  • Perivascular-resident macrophage-like melanocytes
  • Primary cell culture

ASJC Scopus subject areas

  • Sensory Systems

Cite this

Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system. / Zhang, Jinhui; Chen, Songlin; Cai, Jing; Hou, Zhiqiang; Wang, Xiaohan; Kachelmeier, Allan; Shi, Xiao.

In: Hearing Research, 27.06.2016.

Research output: Contribution to journalArticle

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AU - Shi, Xiao

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