Novel Technique for Isolating Human Bone Marrow Stem Cells Using Hyaluronic Acid Hydrogel

Jung Seok Lee, Seul Ki Kim, Jae Kook Cha, Byung Joo Jung, Seong Bok Choi, Eun Young Choi, ChangSung Kim

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Centrifugation based on density gradients is a general methodology for isolating human bone marrow (hBM)-derived mesenchymal stem cells (hBMSCs). The mononuclear cell (MNC) layer can be obtained using a density gradient solution in the conventional protocol, but it is not suitable for direct transplantation due to the possible toxicity of this solution. The results obtained are also influenced by the skill level when applying the technique, which involves time-consuming processes. We have developed a novel protocol for isolating hBMSCs using hyaluronic acid (HA), which is the most widely used injectable biomaterial in clinical settings and a major component of the extracellular matrix. Laying hBM over the HA and then applying centrifugation yielded three separate layers, with the HA layer, including MNCs being the most superficial one. Increasing the volume of HA and/or its crosslinking rate enhanced the yield of MNCs from hBM, and the cell yield was also significantly higher for a lower centrifugal acceleration (530 g) than for a higher one (1500 g). Isolated hBMSCs by HA exhibited similar biological characteristics such as in terms of their proliferation rate, fibroblast-like morphology, cell-cycle status, immunophenotype, and multipotency. The use of either type of hBMSC confirmed the regenerative potential of bone and bone marrow-like tissue in ectopic transplantation models. This is the first report of a novel protocol for isolating hBMSCs that utilize HA. We suggest that this novel isolation technique can be used for the direct application of autogenous MSCs with advantages of being less time-consuming and involving steps that are easier to perform.

Original languageEnglish
Pages (from-to)941-951
Number of pages11
JournalTissue Engineering - Part C: Methods
Volume22
Issue number10
DOIs
Publication statusPublished - 2016 Oct 1

Fingerprint

Hyaluronic acid
Hydrogel
Hyaluronic Acid
Stem cells
Hydrogels
Bone Marrow Cells
Mesenchymal Stromal Cells
Bone
Stem Cells
Centrifugation
Bone Marrow
Transplantation
Cells
Choristoma
Biocompatible Materials
Fibroblasts
Biomaterials
Crosslinking
Extracellular Matrix
Toxicity

All Science Journal Classification (ASJC) codes

  • Bioengineering
  • Medicine (miscellaneous)
  • Biomedical Engineering

Cite this

Lee, Jung Seok ; Kim, Seul Ki ; Cha, Jae Kook ; Jung, Byung Joo ; Choi, Seong Bok ; Choi, Eun Young ; Kim, ChangSung. / Novel Technique for Isolating Human Bone Marrow Stem Cells Using Hyaluronic Acid Hydrogel. In: Tissue Engineering - Part C: Methods. 2016 ; Vol. 22, No. 10. pp. 941-951.
@article{3775d27786e64beab4cde12600d136cf,
title = "Novel Technique for Isolating Human Bone Marrow Stem Cells Using Hyaluronic Acid Hydrogel",
abstract = "Centrifugation based on density gradients is a general methodology for isolating human bone marrow (hBM)-derived mesenchymal stem cells (hBMSCs). The mononuclear cell (MNC) layer can be obtained using a density gradient solution in the conventional protocol, but it is not suitable for direct transplantation due to the possible toxicity of this solution. The results obtained are also influenced by the skill level when applying the technique, which involves time-consuming processes. We have developed a novel protocol for isolating hBMSCs using hyaluronic acid (HA), which is the most widely used injectable biomaterial in clinical settings and a major component of the extracellular matrix. Laying hBM over the HA and then applying centrifugation yielded three separate layers, with the HA layer, including MNCs being the most superficial one. Increasing the volume of HA and/or its crosslinking rate enhanced the yield of MNCs from hBM, and the cell yield was also significantly higher for a lower centrifugal acceleration (530 g) than for a higher one (1500 g). Isolated hBMSCs by HA exhibited similar biological characteristics such as in terms of their proliferation rate, fibroblast-like morphology, cell-cycle status, immunophenotype, and multipotency. The use of either type of hBMSC confirmed the regenerative potential of bone and bone marrow-like tissue in ectopic transplantation models. This is the first report of a novel protocol for isolating hBMSCs that utilize HA. We suggest that this novel isolation technique can be used for the direct application of autogenous MSCs with advantages of being less time-consuming and involving steps that are easier to perform.",
author = "Lee, {Jung Seok} and Kim, {Seul Ki} and Cha, {Jae Kook} and Jung, {Byung Joo} and Choi, {Seong Bok} and Choi, {Eun Young} and ChangSung Kim",
year = "2016",
month = "10",
day = "1",
doi = "10.1089/ten.tec.2016.0214",
language = "English",
volume = "22",
pages = "941--951",
journal = "Tissue Engineering - Part C: Methods",
issn = "1937-3384",
publisher = "Mary Ann Liebert Inc.",
number = "10",

}

Novel Technique for Isolating Human Bone Marrow Stem Cells Using Hyaluronic Acid Hydrogel. / Lee, Jung Seok; Kim, Seul Ki; Cha, Jae Kook; Jung, Byung Joo; Choi, Seong Bok; Choi, Eun Young; Kim, ChangSung.

In: Tissue Engineering - Part C: Methods, Vol. 22, No. 10, 01.10.2016, p. 941-951.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Novel Technique for Isolating Human Bone Marrow Stem Cells Using Hyaluronic Acid Hydrogel

AU - Lee, Jung Seok

AU - Kim, Seul Ki

AU - Cha, Jae Kook

AU - Jung, Byung Joo

AU - Choi, Seong Bok

AU - Choi, Eun Young

AU - Kim, ChangSung

PY - 2016/10/1

Y1 - 2016/10/1

N2 - Centrifugation based on density gradients is a general methodology for isolating human bone marrow (hBM)-derived mesenchymal stem cells (hBMSCs). The mononuclear cell (MNC) layer can be obtained using a density gradient solution in the conventional protocol, but it is not suitable for direct transplantation due to the possible toxicity of this solution. The results obtained are also influenced by the skill level when applying the technique, which involves time-consuming processes. We have developed a novel protocol for isolating hBMSCs using hyaluronic acid (HA), which is the most widely used injectable biomaterial in clinical settings and a major component of the extracellular matrix. Laying hBM over the HA and then applying centrifugation yielded three separate layers, with the HA layer, including MNCs being the most superficial one. Increasing the volume of HA and/or its crosslinking rate enhanced the yield of MNCs from hBM, and the cell yield was also significantly higher for a lower centrifugal acceleration (530 g) than for a higher one (1500 g). Isolated hBMSCs by HA exhibited similar biological characteristics such as in terms of their proliferation rate, fibroblast-like morphology, cell-cycle status, immunophenotype, and multipotency. The use of either type of hBMSC confirmed the regenerative potential of bone and bone marrow-like tissue in ectopic transplantation models. This is the first report of a novel protocol for isolating hBMSCs that utilize HA. We suggest that this novel isolation technique can be used for the direct application of autogenous MSCs with advantages of being less time-consuming and involving steps that are easier to perform.

AB - Centrifugation based on density gradients is a general methodology for isolating human bone marrow (hBM)-derived mesenchymal stem cells (hBMSCs). The mononuclear cell (MNC) layer can be obtained using a density gradient solution in the conventional protocol, but it is not suitable for direct transplantation due to the possible toxicity of this solution. The results obtained are also influenced by the skill level when applying the technique, which involves time-consuming processes. We have developed a novel protocol for isolating hBMSCs using hyaluronic acid (HA), which is the most widely used injectable biomaterial in clinical settings and a major component of the extracellular matrix. Laying hBM over the HA and then applying centrifugation yielded three separate layers, with the HA layer, including MNCs being the most superficial one. Increasing the volume of HA and/or its crosslinking rate enhanced the yield of MNCs from hBM, and the cell yield was also significantly higher for a lower centrifugal acceleration (530 g) than for a higher one (1500 g). Isolated hBMSCs by HA exhibited similar biological characteristics such as in terms of their proliferation rate, fibroblast-like morphology, cell-cycle status, immunophenotype, and multipotency. The use of either type of hBMSC confirmed the regenerative potential of bone and bone marrow-like tissue in ectopic transplantation models. This is the first report of a novel protocol for isolating hBMSCs that utilize HA. We suggest that this novel isolation technique can be used for the direct application of autogenous MSCs with advantages of being less time-consuming and involving steps that are easier to perform.

UR - http://www.scopus.com/inward/record.url?scp=84992694539&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84992694539&partnerID=8YFLogxK

U2 - 10.1089/ten.tec.2016.0214

DO - 10.1089/ten.tec.2016.0214

M3 - Article

C2 - 27609497

AN - SCOPUS:84992694539

VL - 22

SP - 941

EP - 951

JO - Tissue Engineering - Part C: Methods

JF - Tissue Engineering - Part C: Methods

SN - 1937-3384

IS - 10

ER -