潜在候选细胞构建组织工程的泪道上皮: 兔的组织学和细胞学研究

Abstract

Abstract:Objective: Injury and deficiency of the lacrimal duct epithelium (LDE) can lead to a variety of lacrimal diseases. The purpose of this study was to characterize potential candidate cells for constructing a tissue-engineered LDE. Methods: Different areas of the conjunctiva and lacrimal duct tissue were removed from male adult New Zealand white rabbits for histological evaluation. Hematoxylin and eosin staining and immunohistochemical staining of cytokeratin AE1+AE3, cytokeratin 4, Ki-67, and MUC5AC were observed by light microscopy. The surface morphologies of different epithelial tissues and cellular structures were examined using field-emission scanning electron microscopy and transmission electron microscopy. Epithelial cells were isolated from tissues and identified by specific markers. In vitro, proliferative ability and Western blot analyses of the proliferating cell nuclear antigen (PCNA) of different epithelial cells cultured in identical environments were investigated and compared. Results: Histologically, the epithelial specific markers, cytokeratin AE1+AE3 and cytokeratin 4, were expressed in the conjunctiva epithelium and the LDE. Notably, highly proliferative cells stained with Ki-67 were concentrated under the epithelium in a dome structure of the posterior palpebral conjunctiva. Differentiated goblet cells were also found to a lesser extent in this region. Primary palpebral and fornical conjunctival epithelial cells (PFCECs), bulbar conjunctival epithelial cells (BCECs), and lacrimal duct epithelial cells (LDECs) were successfully separated from tissues. In vitro, rabbit PFCECs and LDECs grew faster and expressed more PCNA than BCECs. Conclusions: PFCECs are anatomically similar to LDECs. They also have similar morphological characteristics, immune phenotypes, and proliferation features. PFCECs are therefore potential candidate cells to replace LDECs in tissue engineering to treat lacrimal duct diseases.

A New Isolation Method of Human Lacrimal Canaliculus Epithelial Stem Cells by Maintaining Close Association with Their Niche Cells

“…This forces us to search for other alternatives. One such alternative can be found within the human lacrimal drainage system, consisting of lacrimal puncta, lacrimal canaliculi, common canaliculus, lacrimal sac, and nasolacrimal duct, which plays an important role in draining extra tear fluid into the inferior meatus of the nose 17 , 41 . Our data in this study showed that most of the lacrimal canaliculi had normal epithelial layers (Fig.…”

Role of stem cells in regenerative treatment of dry eye disease caused by lacrimal gland dysfunction

“…LGSCs (Abughanam et al, 2019;Ali et al, 2017;Xiao & Zhang, 2020;Xie et al, 2015;Zoukhri et al, 2008) and side population cells (Mishima et al, 2012). One study (Jeong et al, 2021) evaluated the SC component in LG organoids cultured in vitro (Tables 2 and 3).…”

“…The remaining 25 studies included in this review investigated the development of new drug targets and therapies or focused on direct LG regeneration using SCs from various sources (Table 1). Studies were published between June 2008 (Zoukhri et al, 2008) and September 2020 (Yu et al, 2020) in the United States (Ali et al, 2017; Aluri et al, 2017; Basova et al, 2017; Gromova et al, 2017; Lu et al, 2017; You et al, 2012; Zoukhri et al, 2008), China (Liu, 2017; Xiao & Zhang, 2020; Xie et al, 2015; Yu et al, 2020; Zhang et al, 2014), Germany (Dietrich, Ott, et al, 2019; Dietrich, Roth, et al, 2019; Roth et al, 2015), Canada (Abughanam et al, 2019; Yao et al, 2019), Japan (Hirayama et al, 2013; Mishima et al, 2012), Denmark (Moller‐Hansen et al, 2020), Korea (Jeong et al, 2021; Lee et al, 2015), Brazil (Bittencourt et al, 2016), Spain (Villatoro et al, 2015) and Turkey (Beyazyildiz et al, 2014) (Figure 4).…”

“…Investigated cell types included MSCs (Dietrich, Roth, et al, 2019; Roth et al, 2015; You et al, 2012), ductal cells (Abughanam et al, 2019), acinar cells (Abughanam et al, 2019; Xiao & Zhang, 2020; Zoukhri et al, 2008), LGSCs (Abughanam et al, 2019; Ali et al, 2017; Xiao & Zhang, 2020; Xie et al, 2015; Zoukhri et al, 2008) and side population cells (Mishima et al, 2012). One study (Jeong et al, 2021) evaluated the SC component in LG organoids cultured in vitro (Tables 2 and 3).…”

“…Culture conditions can influence the quality and character of isolated cells, the phenotype of SCs and control differentiation status. The majority of studies used the explant culture method (Dietrich, Roth, et al, 2019; Lu et al, 2017; Xiao & Zhang, 2020; Xie et al, 2015; Zhang et al, 2014) in combination with flow cytometry (Dietrich, Roth, et al, 2019) cell strainers (Dietrich, Roth, et al, 2019) or Matrigel (Lu et al, 2017; Xiao & Zhang, 2020) for cell isolation. Two studies aimed to optimize the culture method specific for cell type: LGSCs (Xiao & Zhang, 2020) and MSCs (Dietrich, Roth, et al, 2019).…”