BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation

Alasdair Gawain Kay; Tina Patricia Dale; Khondoker Mehedi Akram; Param Mohan; Karen Hampson; Nicola Maffulli; Monica A Spiteri; Alicia Jennifer El Haj; Nicholas Robert Forsyth

doi:10.2217/rme.14.67

BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation

Alasdair Gawain Kay
, Tina Patricia Dale
, Khondoker Mehedi Akram
, Param Mohan
, Karen Hampson
, Nicola Maffulli
, Monica A Spiteri
, Alicia Jennifer El Haj
, Nicholas Robert Forsyth

Chemical Engineering

Research output: Contribution to journal › Article › peer-review

11 Citations (Scopus)

Abstract

AIM: Human mesenchymal stem cells (hMSC) are multipotent progenitor cells. We propose the optimization of hMSC isolation and recovery using the application of a controlled hypoxic environment.

MATERIALS & METHODS: We evaluated oxygen, glucose and serum in the recovery of hMSC from bone marrow (BMhMSC). Colony forming units-fibroblastic, cell numbers, tri-lineage differentiation, immunofluorescence and microarray were used to confirm and characterize BMhMSC.

RESULTS: In an optimized (2% O₍₂₎, 4.5 g/l glucose and 5% serum) environment both colony forming units-fibroblastic (p = 0.01) and cell numbers (p = 0.0001) were enhanced over standard conditions. Transcriptional analysis identified differential expression of bone morphogenetic protein 2 (BMP2) and, putatively, chemokine (C-X-C motif) receptor 2 (CXCR2) signaling pathways.

CONCLUSION: We have detailed a potential milestone in the process of refinement of the BMhMSC isolation process.

Original language	English
Pages (from-to)	109-125
Number of pages	17
Journal	Regenerative Medicine
Volume	10
Issue number	2
DOIs	https://doi.org/10.2217/rme.14.67
Publication status	Published - Mar 2015

Keywords

Bone Marrow/pathology
Bone Marrow Cells/cytology
Bone Morphogenetic Protein 2/metabolism
Cell Culture Techniques
Cell Differentiation
Cell Hypoxia
Cell Proliferation
Cells, Cultured
Chemokines/metabolism
Colony-Forming Units Assay
Computational Biology/methods
Glucose/chemistry
Humans
Immunophenotyping
Mesenchymal Stromal Cells/cytology
Microscopy, Fluorescence
Oligonucleotide Array Sequence Analysis
Osteoblasts/cytology
Oxygen/chemistry
Signal Transduction
Transcriptome
Up-Regulation
BMP2
CFU-F
hypoxia
mesenchymal stem cells
physiological normoxia

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Cite this

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title = "BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation",

abstract = "AIM: Human mesenchymal stem cells (hMSC) are multipotent progenitor cells. We propose the optimization of hMSC isolation and recovery using the application of a controlled hypoxic environment.MATERIALS \& METHODS: We evaluated oxygen, glucose and serum in the recovery of hMSC from bone marrow (BMhMSC). Colony forming units-fibroblastic, cell numbers, tri-lineage differentiation, immunofluorescence and microarray were used to confirm and characterize BMhMSC.RESULTS: In an optimized (2\% O(2), 4.5 g/l glucose and 5\% serum) environment both colony forming units-fibroblastic (p = 0.01) and cell numbers (p = 0.0001) were enhanced over standard conditions. Transcriptional analysis identified differential expression of bone morphogenetic protein 2 (BMP2) and, putatively, chemokine (C-X-C motif) receptor 2 (CXCR2) signaling pathways.CONCLUSION: We have detailed a potential milestone in the process of refinement of the BMhMSC isolation process.",

keywords = "Bone Marrow/pathology, Bone Marrow Cells/cytology, Bone Morphogenetic Protein 2/metabolism, Cell Culture Techniques, Cell Differentiation, Cell Hypoxia, Cell Proliferation, Cells, Cultured, Chemokines/metabolism, Colony-Forming Units Assay, Computational Biology/methods, Glucose/chemistry, Humans, Immunophenotyping, Mesenchymal Stromal Cells/cytology, Microscopy, Fluorescence, Oligonucleotide Array Sequence Analysis, Osteoblasts/cytology, Oxygen/chemistry, Signal Transduction, Transcriptome, Up-Regulation, BMP2, CFU-F, hypoxia, mesenchymal stem cells, physiological normoxia",

author = "Kay, \{Alasdair Gawain\} and Dale, \{Tina Patricia\} and Akram, \{Khondoker Mehedi\} and Param Mohan and Karen Hampson and Nicola Maffulli and Spiteri, \{Monica A\} and \{El Haj\}, \{Alicia Jennifer\} and Forsyth, \{Nicholas Robert\}",

year = "2015",

month = mar,

doi = "10.2217/rme.14.67",

language = "English",

volume = "10",

pages = "109--125",

journal = "Regenerative Medicine ",

issn = "1746-0751",

publisher = "Future Medicine",

number = "2",

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TY - JOUR

T1 - BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation

AU - Kay, Alasdair Gawain

AU - Dale, Tina Patricia

AU - Akram, Khondoker Mehedi

AU - Mohan, Param

AU - Hampson, Karen

AU - Maffulli, Nicola

AU - Spiteri, Monica A

AU - El Haj, Alicia Jennifer

AU - Forsyth, Nicholas Robert

PY - 2015/3

Y1 - 2015/3

N2 - AIM: Human mesenchymal stem cells (hMSC) are multipotent progenitor cells. We propose the optimization of hMSC isolation and recovery using the application of a controlled hypoxic environment.MATERIALS & METHODS: We evaluated oxygen, glucose and serum in the recovery of hMSC from bone marrow (BMhMSC). Colony forming units-fibroblastic, cell numbers, tri-lineage differentiation, immunofluorescence and microarray were used to confirm and characterize BMhMSC.RESULTS: In an optimized (2% O(2), 4.5 g/l glucose and 5% serum) environment both colony forming units-fibroblastic (p = 0.01) and cell numbers (p = 0.0001) were enhanced over standard conditions. Transcriptional analysis identified differential expression of bone morphogenetic protein 2 (BMP2) and, putatively, chemokine (C-X-C motif) receptor 2 (CXCR2) signaling pathways.CONCLUSION: We have detailed a potential milestone in the process of refinement of the BMhMSC isolation process.

AB - AIM: Human mesenchymal stem cells (hMSC) are multipotent progenitor cells. We propose the optimization of hMSC isolation and recovery using the application of a controlled hypoxic environment.MATERIALS & METHODS: We evaluated oxygen, glucose and serum in the recovery of hMSC from bone marrow (BMhMSC). Colony forming units-fibroblastic, cell numbers, tri-lineage differentiation, immunofluorescence and microarray were used to confirm and characterize BMhMSC.RESULTS: In an optimized (2% O(2), 4.5 g/l glucose and 5% serum) environment both colony forming units-fibroblastic (p = 0.01) and cell numbers (p = 0.0001) were enhanced over standard conditions. Transcriptional analysis identified differential expression of bone morphogenetic protein 2 (BMP2) and, putatively, chemokine (C-X-C motif) receptor 2 (CXCR2) signaling pathways.CONCLUSION: We have detailed a potential milestone in the process of refinement of the BMhMSC isolation process.

KW - Bone Marrow/pathology

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KW - Bone Morphogenetic Protein 2/metabolism

KW - Cell Culture Techniques

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KW - Cells, Cultured

KW - Chemokines/metabolism

KW - Colony-Forming Units Assay

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KW - Immunophenotyping

KW - Mesenchymal Stromal Cells/cytology

KW - Microscopy, Fluorescence

KW - Oligonucleotide Array Sequence Analysis

KW - Osteoblasts/cytology

KW - Oxygen/chemistry

KW - Signal Transduction

KW - Transcriptome

KW - Up-Regulation

KW - BMP2

KW - CFU-F

KW - hypoxia

KW - mesenchymal stem cells

KW - physiological normoxia

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DO - 10.2217/rme.14.67

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SN - 1746-0751

VL - 10

SP - 109

EP - 125

JO - Regenerative Medicine

JF - Regenerative Medicine

IS - 2

ER -

BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation

Abstract

Keywords

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