Low-frequency, low-magnitude vibrations (LFLM) enhances chondrogenic differentiation potential of human adipose derived mesenchymal stromal stem cells (hASCs)

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Introduction

Materials and Methods

Description of the cell vibration generator prototype

Vibration loading protocol of hASCs culture

Isolation of human adipose derived mesenchymal stem cells (hASCs)

Immunophenotyping, Fluorescence-activated cell sorting (FACs) analysis, and multipotency test

Cell culture

Cell proliferation assay

Examination of hASCs morphology

Enzyme-linked immunosorbent assays

Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR)

Statistical analysis

Results

hASCs—FACs analysis, immunophenotyping and multipotency test

Proliferation rate (PF) and population doubling time (PDT) of 3D chondroblasts originated from chondro induced hASCs

The morphology of chondroblasts originated from hASCs

Quantitative Collagen 1 and 2 (Col-1, Col-2) and Bone Morphogenetic Protein 2 (BMP-2) assay and chondrogenic gene expression analysis (SOX-9, Col-X, Col-II, Runx, Col-I, ACAN)

Analysis of integrin expression in response to vibration stimulation

Proliferation factor and population doubling time of human adipocytes originated from adipo induced hASCs

Morphology of ASCs after adipogenic differentiation

Discussion

Conclusions

Supplemental Information

SOX9 qPCR results

DOI: 10.7717/peerj.1637/supp-1

collagen X q pcr results

DOI: 10.7717/peerj.1637/supp-2

Col-2 qPCR results

DOI: 10.7717/peerj.1637/supp-3

RUNX qPCR results

DOI: 10.7717/peerj.1637/supp-4

Col-1 qPCR results

DOI: 10.7717/peerj.1637/supp-5

Aggrecan qPCR results

DOI: 10.7717/peerj.1637/supp-6

Col-1 ELISA results

DOI: 10.7717/peerj.1637/supp-7

BMP-2 ELISA results

DOI: 10.7717/peerj.1637/supp-8

Col-2 ELISA results

DOI: 10.7717/peerj.1637/supp-9

Integrin alfa 3 qPCR results

DOI: 10.7717/peerj.1637/supp-10

Integrin alfa 4 qPCR results

DOI: 10.7717/peerj.1637/supp-11

Integrin alfa 5 qPCR results

DOI: 10.7717/peerj.1637/supp-12

Integrin alfa V qPCR results

DOI: 10.7717/peerj.1637/supp-13

Integrin Beta 1 qPCR results

DOI: 10.7717/peerj.1637/supp-14

Integrin beta 3 qPCR results

DOI: 10.7717/peerj.1637/supp-15

Integrin beta 5 qPCR results

DOI: 10.7717/peerj.1637/supp-16

PPAR-gamma qPCR results

DOI: 10.7717/peerj.1637/supp-17

Adiponectin qPCR results

DOI: 10.7717/peerj.1637/supp-18

LEPTIN qPCR results

DOI: 10.7717/peerj.1637/supp-19

Average diameters of nodules

DOI: 10.7717/peerj.1637/supp-20

proliferation factor

DOI: 10.7717/peerj.1637/supp-21

Population doubling time

DOI: 10.7717/peerj.1637/supp-22

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests regarding the publication of this paper.

Author Contributions

Krzysztof Marycz conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, reviewed drafts of the paper.

Daniel Lewandowski conceived and designed the experiments, performed the experiments, wrote the paper.

Krzysztof A. Tomaszewski analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, reviewed drafts of the paper.

Brandon Michael Henry analyzed the data, wrote the paper, reviewed drafts of the paper.

Edward B. Golec contributed reagents/materials/analysis tools, reviewed drafts of the paper.

Monika Marędziak performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables.

Human Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

The study was approved by the local bioethics committee of Wroclaw Medical University, Poland (number KB-177/2014). Written informed consent was obtained from each patient prior to tissue collection during total hip arthroplasty. This study adhered to the Helsinki Declaration (1964) and its later amendments.

Data Availability

The following information was supplied regarding data availability:

The raw data is provided as Supplemental Dataset files. The results were generated via GraphPad Prism and exported to csv files.

Funding

The research was supported by Wroclaw Research Centre EIT + under the project ‘Biotechnologies and advanced medical technologies’—BioMed (POIG.01.01.02-02-003/08) financed from the European Regional Development Fund (Operational Programmed Innovative Economy, 1.1.2.). This publication was supported by Wrocław Centre of Biotechnology, the Leading National Research Centre (KNOW) program between 2014 and 2018. Krzysztof A. Tomaszewski was supported by the Foundation for Polish Science (FNP). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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