CC BY-NC-ND 4.0 · Homeopathy
DOI: 10.1055/s-0042-1747682
Original Research Article

In-Vitro Growth Kinetics of Mesenchymal Stem Cells in Cytotoxicity Tests Using Low-Diluted Viscum Album

Ana Catarina Viana Valle
1   Doctor Izao Soares Institute, Sao Paulo, Brazil
2   Graduate Program in Genomic Sciences and Biotechnology, Catholic University of Brasilia, Brazil
,
Hilana dos Santos Sena Brunel
3   BioInnova, Brazil
,
Bruno Stéfano Lima Dallago
4   Brasilia University, Brazil
,
Lucas Santana Rodrigues
5   BioCell Cell Therapy, Brazil
,
Patrícia Furtado Malard
2   Graduate Program in Genomic Sciences and Biotechnology, Catholic University of Brasilia, Brazil
5   BioCell Cell Therapy, Brazil
,
Rosiane Andrade da Costa
2   Graduate Program in Genomic Sciences and Biotechnology, Catholic University of Brasilia, Brazil
,
Rafael Rossetto
2   Graduate Program in Genomic Sciences and Biotechnology, Catholic University of Brasilia, Brazil
,
2   Graduate Program in Genomic Sciences and Biotechnology, Catholic University of Brasilia, Brazil
› Author Affiliations
Funding This research and the manuscript preparation did not receive any funding. RAC received a post-doctoral scholarship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/PNPD protocol 88882.315167/2019–01).

Abstract

Introduction The use of mesenchymal stem cells (MSC) in cytotoxicity tests is an in-vitro alternative model for predicting initial doses. Homeopathic medicines may stimulate the immune system to combat a pathology effectively and have been used for over two centuries. Viscum album (VA) extracts are widely used in the treatment of cancer, due to their immunomodulatory, cytotoxic and pro-apoptotic properties.

Objective This study aimed to evaluate the in-vitro growth kinetics of canine MSC in relation to cytotoxicity, cell differentiation and expression of pluripotentiality markers, using a VA preparation at the D1D2 (1×10−1, 1×10−2 potency (VAD1D2).

Methods MSC were obtained from adipose tissue sampled from a healthy dog that was undergoing an elective veterinary procedure and with its owner's permission. The experiments were performed in three groups: MSC treated with VAD1D2 or diluent or untreated (control). The cytotoxicity was evaluated by MTT assay. The differentiation was induced in three lineages, and apoptotic cell labeling was performed by an Annexin-V test.

Results At the concentration of 10 μL/mL of VA, the number of cells after in-vitro culture was maintained when compared with the control (untreated) group. A significant and gradual decrease in cell viability was recorded as VA concentrations increased. The apoptosis analysis showed that VA at 20 μL/mL presented absolute percentages of initial apoptosis twice as high as at 10 μL/mL, which was similar to the control (untreated group).

Conclusion The results suggest that the use of efficient methods to assess the in-vitro cytotoxicity of VA-based homeopathic medicines using MSC lineages may predict the potential action at different concentrations. These findings demonstrated that VAD1D2 interferes with canine MSC growth kinetics.

Authors' Contributions

A.C.V.V. conceptualized the study, and contributed to the methodology, investigation, formal analysis, and writing—original draft H.d.S.S.B. contributed to methodology, investigation, formal analysis, and writing—original draft. B.S.L.D. did the formal analysis and co-wrote the original draft. L.S.R. contributed to conceptualization, investigation, and acquisition of resources. P.F.M. contributed to conceptualization and investigation, formal analysis, and writing—original draft. R.R. contributed to conceptualization, writing—review and editing. R.A.C. contributed to co-writing—review and editing. R.V.A. contributed to conceptualization, methodology, supervision, and project administration. All authors critically reviewed the manuscript, contributed to its revision, and approved the final version submitted.


Supplementary Material



Publication History

Received: 15 October 2021

Accepted: 14 January 2022

Article published online:
21 August 2022

© 2022. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany