Ravindra K. Zirmire
  • Centre for Inflammation and Tissue Homeostasis, Institute for Stem Cell Science and Regenerative Medicine, Bangalore
Isolation and Quantification of Mouse γδT-cells in vitro and in vivo
体外和体内小鼠γδT细胞的分离和定量
作者:Isha Rana, Krithika Badarinath, Ravindra K. Zirmire and Colin Jamora日期:09/05/2021,浏览量:3101,Q&A: 0

The skin plays an important role in protecting the body from pathogens and chemicals in the external environment. Upon injury, a healing program is rapidly initiated and involves extensive intercellular communication to restore tissue homeostasis. The deregulation of this crosstalk can lead to abnormal healing processes and is the foundation of many skin diseases. A relatively overlooked cell type that nevertheless plays critical roles in skin homeostasis, wound repair, and disease is the dendritic epidermal T cells (DETCs), which are also called γδT-cells. Given their varied roles in both physiological and pathological scenarios, interest in the regulation and function of DETCs has substantially increased. Moreover, their ability to regulate other immune cells has garnered substantial attention for their potential role as immunomodulators and in immunotherapies. In this article, we describe a protocol to isolate and culture DETCs and analyse them in vivo within the skin. These approaches will facilitate the investigation of their crosstalk with other cutaneous cells and the mechanisms by which they influence the status of the skin.



Graphic abstract:



Overall workflow to analyse DETCs in vitro and in vivo.


Histological and Immunohistochemical Examination of Stem Cell Proliferation and Reepithelialization in the Wounded Skin
损伤皮肤干细胞增殖和再生的组织学和免疫组化检测
作者:Rupali Gund, Ravindra Zirmire, Haarshaadri J., Gaurav Kansagara and Colin Jamora日期:01/20/2021,浏览量:4787,Q&A: 0

The skin is the largest organ that protects our body from the external environment and it is constantly exposed to pathogenic insults and injury. Repair of damage to this organ is carried out by a complex process involving three overlapping phases of inflammation, proliferation and remodeling. Histological analysis of wounded skin is a convenient approach to examine broad alterations in tissue architecture and investigate cells in their indigenous microenvironment. In this article we present a protocol for immunohistochemical examination of wounded skin to study mechanisms involved in regulating stem cell activity, which is a vital component in the repair of the damaged tissue. Performing such histological analysis enables the understanding of the spatial relationship between cells that interact in the specialized wound microenvironment. The analytical tools described herein permit the quantitative measurement of the regenerative ability of stem cells adjacent to the wound and the extent of re-epithelialization during wound closure. These protocols can be adapted to investigate numerous cellular processes and cell types within the wounded skin.

Activation of Fibroblast Contractility via Cell-Cell Interactions and Soluble Signals
通过细胞间相互作用和可溶信号激活成纤维细胞收缩
作者:Neha Pincha, Dyuti Saha, Tanay Bhatt, Ravindra K. Zirmire and Colin Jamora日期:09/20/2018,浏览量:8159,Q&A: 0
The collagen contraction assay is an in vitro, three-dimensional method to determine the factor(s) affecting the contractile behavior of activated cells such as fibroblasts in either physiological or pathological scenarios. The collagen lattices/hydrogels are seeded with fibroblasts to mimic the interactions between these cells and their surrounding extracellular matrix proteins in the connective tissue. This method is an important platform to assess components as potential therapeutic targets to prevent pathologies such as fibrosis, which are manifestations of hyperactivated fibroblasts. We have described a basic version of this collagen contraction assay, which is amenable to customization using different cell types under diverse experimental conditions.