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Methods and Protocols, Volume 5, Issue 1

February 2022 - 19 articles

Cover Story: New biomaterials for bone tissue engineering (BTE) applications need to be tested in a reliable model of bone microenvironment, and in vitro tests with bone cells represent worthy systems. Herein, a clear protocol to set up an indirect co-culture system of human-derived osteoblasts (OBs) and osteoclasts (OCs), reporting cell seeding density, cell:cell ratio, culture medium, and proofs of differentiation, is provided. The material to be tested is easily introduced in the system and the physical separation of OBs and OCs allows distinguishing the effects of the material on the two cell types via direct/indirect interaction between material and cells. The system is an in vitro model of bone coupling, useful in testing innovative materials for bone regeneration, and potentially exploitable in different application fields. View this paper
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Articles (19)

  • Protocol
  • Open Access
4 Citations
4,040 Views
15 Pages

16 February 2022

Members of the saposin-fold protein family and related proteins sharing a similar fold (saposin-like proteins; SAPLIP) are peripheral-membrane binding proteins that perform essential cellular functions. Saposins and SAPLIPs are abundant in both plant...

  • Article
  • Open Access
1 Citations
3,869 Views
13 Pages

Harnessing Intronic microRNA Structures to Improve Tolerance and Expression of shRNAs in Animal Cells

  • Arjun Challagulla,
  • Mark L. Tizard,
  • Timothy J. Doran,
  • David M. Cahill and
  • Kristie A. Jenkins

10 February 2022

Exogenous RNA polymerase III (pol III) promoters are commonly used to express short hairpin RNA (shRNA). Previous studies have indicated that expression of shRNAs using standard pol III promoters can cause toxicity in vivo due to saturation of the na...

  • Review
  • Open Access
49 Citations
18,669 Views
13 Pages

9 February 2022

Cell monolayers, including endothelial and epithelial cells, play crucial roles in regulating the transport of biomolecules to underlying tissues and structures via intercellular junctions. Moreover, the monolayers form a semipermeable barrier across...

  • Protocol
  • Open Access
7 Citations
3,550 Views
9 Pages

Development of Primary Monolayer Cell Model and Organotypic Model of Uterine Leiomyoma

  • Natalia Shved,
  • Anna Egorova,
  • Natalia Osinovskaya and
  • Anton Kiselev

6 February 2022

Cellular technologies are one of the most promising areas of biomedicine, which is based on the isolation of cells of various types, followed by their cultivation and use, or the use of their metabolic products, for medical purposes. Today, a signifi...

  • Communication
  • Open Access
6 Citations
3,411 Views
9 Pages

5 February 2022

Proteasomes hydrolyze most cellular proteins. The standard reaction to determine proteasome activity in cellular lysate or elsewhere contains AMC-conjugated peptide substrate, ATP, Mg2+, and DTT. ATP and Mg2+ are included to maintain 26S proteasome f...

  • Article
  • Open Access
9 Citations
5,184 Views
25 Pages

Knotify: An Efficient Parallel Platform for RNA Pseudoknot Prediction Using Syntactic Pattern Recognition

  • Christos Andrikos,
  • Evangelos Makris,
  • Angelos Kolaitis,
  • Georgios Rassias,
  • Christos Pavlatos and
  • Panayiotis Tsanakas

2 February 2022

Obtaining valuable clues for noncoding RNA (ribonucleic acid) subsequences remains a significant challenge, acknowledging that most of the human genome transcribes into noncoding RNA parts related to unknown biological operations. Capturing these clu...

  • Protocol
  • Open Access
17 Citations
6,836 Views
7 Pages

A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections

  • Divya Sridharan,
  • Nooruddin Pracha,
  • Julie A. Dougherty,
  • Ali Akhtar,
  • Syed Baseeruddin Alvi and
  • Mahmood Khan

27 January 2022

Masson’s Trichrome Staining (MTS) is a useful tool for analyzing fibrosis in a plethora of disease pathologies by differential staining of tissue components. It is used to identify collagen fibers in different tissues like heart, lung, skin, an...

  • Article
  • Open Access
8 Citations
7,461 Views
17 Pages

A Simple and Effective Bioassay Method Suitable to Comparative In Vitro Study of Tomato Salt Tolerance at Early Development Stages

  • Marat R. Khaliluev,
  • Liliya R. Bogoutdinova,
  • Galina N. Raldugina and
  • Ekaterina N. Baranova

19 January 2022

In vitro evaluation of tomato seeds and seedlings for salt tolerance has undoubted advantages (high productivity, as well as stability and reproducibility of the obtained experimental data due to the maintenance of constant controlled conditions) in...

  • Article
  • Open Access
5 Citations
4,504 Views
14 Pages

PCA-Assisted Raman Analysis of Osteonecrotic Human Femoral Heads

  • Eiji Ishimura,
  • Wenliang Zhu,
  • Elia Marin,
  • Taigi Honma,
  • Nobuhiko Sugano,
  • Wataru Ando and
  • Giuseppe Pezzotti

17 January 2022

Osteonecrosis of the femoral head (ONFH) occurs frequently in adolescents and young adults and causes progressive deformation and destruction of the hip joint and impairs standing and walking, resulting in a significant decrease in the quality of lif...

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Methods Protoc. - ISSN 2409-9279