Human bone marrow-derived MSCs spontaneously express specific Schwann cell markers

Khairunnisa Ramli, Ifasha Aminath Gasim, Amir Adham Ahmad, Shariful Hassan, Zhe Kang Law, Geok Chin Tan, Azmi Baharuddin, Amaramalar Selvi Naicker, Ohnmar Htwe, Nor Hazla Mohammed Haflah, Ruszymah B. H. Idrus, Shalimar Abdullah, Min Hwei Ng

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In peripheral nerve injuries, Schwann cells (SC) play pivotal roles in regenerating damaged nerve. However, the use of SC in clinical cell-based therapy is hampered due to its limited availability. In this study, we aim to evaluate the effectiveness of using an established induction protocol for human bone marrow derived-MSC (hBM-MSCs) transdifferentiation into a SC lineage. A relatively homogenous culture of hBM-MSCs was first established after serial passaging (P3), with profiles conforming to the minimal criteria set by International Society for Cellular Therapy (ISCT). The cultures (n = 3) were then subjected to a series of induction media containing β-mercaptoethanol, retinoic acid, and growth factors. Quantitative RT-PCR, flow cytometry, and immunocytochemistry analyses were performed to quantify the expression of specific SC markers, that is, S100, GFAP, MPZ and p75 NGFR, in both undifferentiated and transdifferentiated hBM-MSCs. Based on these analyses, all markers were expressed in undifferentiated hBM-MSCs and MPZ expression (mRNA transcripts) was consistently detected before and after transdifferentiation across all samples. There was upregulation at the transcript level of more than twofolds for NGF, MPB, GDNF, p75 NGFR post-transdifferentiation. This study highlights the existence of spontaneous expression of specific SC markers in cultured hBM-MSCs, inter-donor variability and that MSC transdifferentiation is a heterogenous process. These findings strongly oppose the use of a single marker to indicate SC fate. The heterogenous nature of MSC may influence the efficiency of SC transdifferentiation protocols. Therefore, there is an urgent need to re-define the MSC subpopulations and revise the minimal criteria for MSC identification.

Original languageEnglish
JournalCell Biology International
DOIs
Publication statusAccepted/In press - 1 Jan 2019

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Schwann Cells
Bone Marrow
Cell Transdifferentiation
Glial Cell Line-Derived Neurotrophic Factor
Peripheral Nerve Injuries
Mercaptoethanol
Nerve Growth Factor
Cell Lineage
Cell- and Tissue-Based Therapy
Tretinoin
Intercellular Signaling Peptides and Proteins
Flow Cytometry
Up-Regulation
Immunohistochemistry
Polymerase Chain Reaction
Messenger RNA

Keywords

  • heterogeneity
  • human mesenchymal stem cells
  • Schwann cells
  • transdifferentiation

ASJC Scopus subject areas

  • Cell Biology

Cite this

Human bone marrow-derived MSCs spontaneously express specific Schwann cell markers. / Ramli, Khairunnisa; Aminath Gasim, Ifasha; Ahmad, Amir Adham; Hassan, Shariful; Law, Zhe Kang; Tan, Geok Chin; Baharuddin, Azmi; Selvi Naicker, Amaramalar; Htwe, Ohnmar; Mohammed Haflah, Nor Hazla; B. H. Idrus, Ruszymah; Abdullah, Shalimar; Ng, Min Hwei.

In: Cell Biology International, 01.01.2019.

Research output: Contribution to journalArticle

Ramli, Khairunnisa ; Aminath Gasim, Ifasha ; Ahmad, Amir Adham ; Hassan, Shariful ; Law, Zhe Kang ; Tan, Geok Chin ; Baharuddin, Azmi ; Selvi Naicker, Amaramalar ; Htwe, Ohnmar ; Mohammed Haflah, Nor Hazla ; B. H. Idrus, Ruszymah ; Abdullah, Shalimar ; Ng, Min Hwei. / Human bone marrow-derived MSCs spontaneously express specific Schwann cell markers. In: Cell Biology International. 2019.
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abstract = "In peripheral nerve injuries, Schwann cells (SC) play pivotal roles in regenerating damaged nerve. However, the use of SC in clinical cell-based therapy is hampered due to its limited availability. In this study, we aim to evaluate the effectiveness of using an established induction protocol for human bone marrow derived-MSC (hBM-MSCs) transdifferentiation into a SC lineage. A relatively homogenous culture of hBM-MSCs was first established after serial passaging (P3), with profiles conforming to the minimal criteria set by International Society for Cellular Therapy (ISCT). The cultures (n = 3) were then subjected to a series of induction media containing β-mercaptoethanol, retinoic acid, and growth factors. Quantitative RT-PCR, flow cytometry, and immunocytochemistry analyses were performed to quantify the expression of specific SC markers, that is, S100, GFAP, MPZ and p75 NGFR, in both undifferentiated and transdifferentiated hBM-MSCs. Based on these analyses, all markers were expressed in undifferentiated hBM-MSCs and MPZ expression (mRNA transcripts) was consistently detected before and after transdifferentiation across all samples. There was upregulation at the transcript level of more than twofolds for NGF, MPB, GDNF, p75 NGFR post-transdifferentiation. This study highlights the existence of spontaneous expression of specific SC markers in cultured hBM-MSCs, inter-donor variability and that MSC transdifferentiation is a heterogenous process. These findings strongly oppose the use of a single marker to indicate SC fate. The heterogenous nature of MSC may influence the efficiency of SC transdifferentiation protocols. Therefore, there is an urgent need to re-define the MSC subpopulations and revise the minimal criteria for MSC identification.",
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AU - Ramli, Khairunnisa

AU - Aminath Gasim, Ifasha

AU - Ahmad, Amir Adham

AU - Hassan, Shariful

AU - Law, Zhe Kang

AU - Tan, Geok Chin

AU - Baharuddin, Azmi

AU - Selvi Naicker, Amaramalar

AU - Htwe, Ohnmar

AU - Mohammed Haflah, Nor Hazla

AU - B. H. Idrus, Ruszymah

AU - Abdullah, Shalimar

AU - Ng, Min Hwei

PY - 2019/1/1

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AB - In peripheral nerve injuries, Schwann cells (SC) play pivotal roles in regenerating damaged nerve. However, the use of SC in clinical cell-based therapy is hampered due to its limited availability. In this study, we aim to evaluate the effectiveness of using an established induction protocol for human bone marrow derived-MSC (hBM-MSCs) transdifferentiation into a SC lineage. A relatively homogenous culture of hBM-MSCs was first established after serial passaging (P3), with profiles conforming to the minimal criteria set by International Society for Cellular Therapy (ISCT). The cultures (n = 3) were then subjected to a series of induction media containing β-mercaptoethanol, retinoic acid, and growth factors. Quantitative RT-PCR, flow cytometry, and immunocytochemistry analyses were performed to quantify the expression of specific SC markers, that is, S100, GFAP, MPZ and p75 NGFR, in both undifferentiated and transdifferentiated hBM-MSCs. Based on these analyses, all markers were expressed in undifferentiated hBM-MSCs and MPZ expression (mRNA transcripts) was consistently detected before and after transdifferentiation across all samples. There was upregulation at the transcript level of more than twofolds for NGF, MPB, GDNF, p75 NGFR post-transdifferentiation. This study highlights the existence of spontaneous expression of specific SC markers in cultured hBM-MSCs, inter-donor variability and that MSC transdifferentiation is a heterogenous process. These findings strongly oppose the use of a single marker to indicate SC fate. The heterogenous nature of MSC may influence the efficiency of SC transdifferentiation protocols. Therefore, there is an urgent need to re-define the MSC subpopulations and revise the minimal criteria for MSC identification.

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