Authors: Prakash Baligar
, Sebanta Pokhrel Prakash BaligarAffiliation:
Amity Institute of Molecular Medicine and Stem Cell Research (AIMMSCR), Amity University, Noida, UP, IndiaFor correspondence: firstname.lastname@example.orgBio-protocol author page: a3620
and Asok Mukhopadhyay Sebanta PokhrelAffiliation:
Stem Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, IndiaBio-protocol author page: a3621
date: 10/20/2016, 23 views, 0 Q&A.
Stem Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, IndiaFor correspondence: email@example.comBio-protocol author page: a3622
|Brief version appeared in Stem Cells, Jan 2016 |
Liver fibrosis results from the excessive collagen deposition (collagen scar) by activated hepatic stellate cells (HpSCs), leading to the inhibition of normal liver regeneration and function. Fibrogenesis is a complex mechanism involving both the synthesis and degradation of matrix proteins by different cell types, mainly macrophages in the liver. Carbon tetrachloride-induced fibrosis (CCl4
) and cirrhosis is one of the oldest, simplest and probably the most widely used toxin-based experimental model for the induction of fibrosis. Here we have explained experimental animal model of liver fibrosis using CCl4
, injecting twice a week for a period of 8 weeks. In these fibrotic mice, bone marrow (BM) derived CD45+
cells were transplanted via intrasplenic route after 8 weeks of CCl4
injection, and half of the CCl4
dose was continued till the end of the experiment to know the effect of transplanted cells on liver fibrosis and regeneration. So far, crude bone marrow (BM) cells or mesenchymal stem cells (MSCs) have been used for the treatment of liver fibrosis. Low survival rate, less fibrolytic and profibrogenic properties of MSCs remain the major concerns for inadequate recovery of liver from fibrosis. This led us to investigate BM cells devoid of mesenchymal lineage that is CD45+
cells for the antifibrotic effect as this population consisting of mononuclear cells which are the precursor of macrophages and may involve in the scar degradation process. Cells transplantation can be followed in different ways like intrasplenic infusion, tail vein injection and ectopic cell transplantation in experimental animal models. The survival of the cells after ectopic transplantation is less when compared to tail vein and intrasplenic infusion. Intrasplenic route of transplantation is effective in engraftment and long term survival of the donor cells especially in case of liver disease models. This protocol describes fibrosis mouse model development, intrasplenic route of cell transplantation and tracking of the donor cells after transplantation.
Authors: Yann Simon Gallot
, Sajedah M. Hindi Yann Simon GallotAffiliation:
Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USABio-protocol author page: a3535
, Aman K. Mann Sajedah M. HindiAffiliation:
Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USABio-protocol author page: a3536
and Ashok Kumar Aman K. MannAffiliation:
duPont Manual High Schoo, Louisville, USABio-protocol author page: a3538
date: 10/5/2016, 75 views, 0 Q&A.
Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USAFor correspondence: firstname.lastname@example.orgBio-protocol author page: a3537
|Brief version appeared in J Clin Invest, Jan 2016 |
Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo
by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate ex vivo
satellite cells activation through isolation of single myofibers from extensor digitorum longus (EDL) muscle of mice and culturing and staining of myofiber-associated satellite cells with the markers of self-renewal, proliferation, and differentiation.