Featured protocol,
Authors: Ji Yong LeeJi Yong LeeAffiliation: Institute for BioMedical Convergence, Catholic Kwandong University-International St. Mary’s Hospital, Incheon-si, Republic of Korea
Bio-protocol author page: a4768 Seong-Mi ChoiAffiliation: Institute for BioMedical Convergence, Catholic Kwandong University-International St. Mary’s Hospital, Incheon-si, Republic of Korea
Bio-protocol author page: a4769 
Han-Soo KimAffiliation: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea
For correspondence: hankim63@gmail.comBio-protocol author page: a4770
DOI: https://doi.org/10.21769/BioProtoc.2375.
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| Brief version appeared in Sci Rep, Sep 2016 |
Microvesicle (MVs) are submicron-sized membranous vesicles that are either actively released from cells via secretory compartments or shed from cell surface membranes. MVs are generated by many cell types and serve as vehicles that transfer biological information (
e.g., protein, mRNA, and miRNA) to distant cells, thereby affecting their gene expression, proliferation, differentiation, and function. Although their physiological functions are not clearly defined, recent studies have shown their therapeutic potential for tissue repair and regeneration. While MVs can be isolated readily from mesenchymal stem cells (MSCs) and other cell types from various sources, the yield of MVs under conventional culture condition
in vitro is one of the limiting factors for both the
in vivo functional study as well as
in vitro molecular analysis. Here, we provide a protocol to increase the yield of microvesicles by preconditioning MSCs with rat brain extract.
Featured protocol,
Authors: Natalia V. AndreevaNatalia V. AndreevaAffiliation: Laboratory of Stem and Progenitor Cell Biology, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia
Bio-protocol author page: a4766 Alexander V. BelyavskyAffiliation: Laboratory of Stem and Progenitor Cell Biology, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia
For correspondence: abelyavs@yahoo.comBio-protocol author page: a4767
DOI: https://doi.org/10.21769/BioProtoc.2374.
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| Brief version appeared in Anal Biochem, Dec 2016 |
Coating tissue culture vessels with the components of the extracellular matrix such as fibronectin and collagens provides a more natural environment for primary cells
in vitro and stimulates their proliferation. However, the effects of such protein layers are usually rather modest, which might be explained by the loss immobilized proteins due to their weak non-covalent association with the tissue culture plastic. Here we describe a simple protocol for a controlled fixation of fibronectin, vitronectin and collagen IV layers by formaldehyde, which substantially enhances the stimulation of primary cell proliferation by these extracellular proteins.
Featured protocol,
Authors: Cong WangCong WangAffiliation: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou, China
Bio-protocol author page: a4734 Hao GaoAffiliation: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou, China
Bio-protocol author page: a4735 Shengxiang ZhangAffiliation: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou, China
For correspondence: sxzhang@lzu.edu.cnBio-protocol author page: a4736
DOI: https://doi.org/10.21769/BioProtoc.2360.
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| Brief version appeared in Sci Rep, Sep 2016 |
Transplantation of embryonic cortical tissue for repairing the damaged brain has provided a potential therapy for brain injury and diseases. The grafted tissue can successfully survive and participate in reestablishing the functional neural circuit of the host brain. Transplantation surgery can be combined with fluorescently labeled transgenic mice to evaluate the reconstruction of neuronal network (Falkner
et al., 2016) and the repopulation of a subset of cortical cells. By using this approach, we have shown that infiltrating cells from host brain can restore the microglial population in the graft tissue (Wang
et al., 2016). This protocol describes the detailed procedure of the transplantation surgery in mice, including establishing a lesion model in the host brain, preparing the embryonic cortical graft, and transplanting the embryonic cortical graft to adult brain.