Human adipose-derived stromal cells in a clinically applicable injectable alginate hydrogel: Phenotypic and immunomodulatory evaluation

Bjarke Follin Larsen; Morten Juhl; Smadar Cohen; Anders Elm Pedersen; Monika Gad; Jens Kastrup; Annette Ekblond

doi:10.1016/j.jcyt.2015.04.008

Human adipose-derived stromal cells in a clinically applicable injectable alginate hydrogel: Phenotypic and immunomodulatory evaluation

Bjarke Follin Larsen, Morten Juhl, Smadar Cohen, Anders Elm Pedersen, Monika Gad, Jens Kastrup, Annette Ekblond

Institut for Klinisk Medicin

30 Citationer (Scopus)

Abstract

BACKGROUND AIMS: Clinical trials have documented beneficial effects of mesenchymal stromal cells from bone marrow and adipose tissue (ASCs) as treatment in patients with ischemic heart disease. However, retention of transplanted cells is poor. One potential way to increase cell retention is to inject the cells in an in situ cross-linked alginate hydrogel.

METHODS: ASCs from abdominal human tissue were embedded in alginate hydrogel and alginate hydrogel modified with Arg-Gly-Asp motifs (RGD-alginate) and cultured for 1 week. Cell viability, phenotype, immunogenicity and paracrine activity were determined by confocal microscopy, dendritic cell co-culture, flow cytometry, reverse transcriptase quantitative polymerase chain reaction, Luminex multiplex, and lymphocyte proliferation experiments.

RESULTS: ASCs performed equally well in alginate and RGD-alginate. After 1 week of alginate culture, cell viability was >93%. Mesenchymal markers CD90 and CD29 were reduced compared with International Society for Cellular Therapy criteria. Cells sedimented from the alginates during cultivation regained the typical level of these markers, and trilineage differentiation was performed by standard protocols. Hepatocyte growth factor mRNA was increased in ASCs cultivated in alginates compared with monolayer controls. Alginates and alginates containing ASCs did not induce dendritic cell maturation. ASCs in alginate responded like controls to interferon-gamma stimulation (licensing), and alginate culture increased the ability of ASCs to inhibit lymphocyte proliferation.

DISCUSSION: ASCs remain viable in alginates; they transiently change phenotype in alginate hydrogel but regain the phenotype of monolayer controls upon release. Cells maintain their paracrine potential while in alginates; the combination of ASCs and alginate is non-immunogenic and, in fact, immunosuppressive.

Originalsprog	Engelsk
Tidsskrift	Cytotherapy
Vol/bind	17
Udgave nummer	8
Sider (fra-til)	1104-18
Antal sider	15
ISSN	1465-3249
DOI	https://doi.org/10.1016/j.jcyt.2015.04.008
Status	Udgivet - 1 aug. 2015

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10.1016/j.jcyt.2015.04.008

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title = "Human adipose-derived stromal cells in a clinically applicable injectable alginate hydrogel: Phenotypic and immunomodulatory evaluation",

abstract = "BACKGROUND AIMS: Clinical trials have documented beneficial effects of mesenchymal stromal cells from bone marrow and adipose tissue (ASCs) as treatment in patients with ischemic heart disease. However, retention of transplanted cells is poor. One potential way to increase cell retention is to inject the cells in an in situ cross-linked alginate hydrogel.METHODS: ASCs from abdominal human tissue were embedded in alginate hydrogel and alginate hydrogel modified with Arg-Gly-Asp motifs (RGD-alginate) and cultured for 1 week. Cell viability, phenotype, immunogenicity and paracrine activity were determined by confocal microscopy, dendritic cell co-culture, flow cytometry, reverse transcriptase quantitative polymerase chain reaction, Luminex multiplex, and lymphocyte proliferation experiments.RESULTS: ASCs performed equally well in alginate and RGD-alginate. After 1 week of alginate culture, cell viability was >93%. Mesenchymal markers CD90 and CD29 were reduced compared with International Society for Cellular Therapy criteria. Cells sedimented from the alginates during cultivation regained the typical level of these markers, and trilineage differentiation was performed by standard protocols. Hepatocyte growth factor mRNA was increased in ASCs cultivated in alginates compared with monolayer controls. Alginates and alginates containing ASCs did not induce dendritic cell maturation. ASCs in alginate responded like controls to interferon-gamma stimulation (licensing), and alginate culture increased the ability of ASCs to inhibit lymphocyte proliferation.DISCUSSION: ASCs remain viable in alginates; they transiently change phenotype in alginate hydrogel but regain the phenotype of monolayer controls upon release. Cells maintain their paracrine potential while in alginates; the combination of ASCs and alginate is non-immunogenic and, in fact, immunosuppressive.",

keywords = "Adipocytes, Adipose Tissue, Adult, Aged, Aged, 80 and over, Alginates, Cell Differentiation, Cell Proliferation, Cell Survival, Cells, Cultured, Coculture Techniques, Female, Glucuronic Acid, Hepatocyte Growth Factor, Hexuronic Acids, Humans, Hydrogel, Immunomodulation, Interferon-gamma, Male, Mesenchymal Stem Cell Transplantation, Mesenchymal Stromal Cells, Middle Aged, Oligopeptides, RNA, Messenger, Tissue Embedding, Young Adult",

author = "Larsen, {Bjarke Follin} and Morten Juhl and Smadar Cohen and Pedersen, {Anders Elm} and Monika Gad and Jens Kastrup and Annette Ekblond",

year = "2015",

month = aug,

day = "1",

doi = "10.1016/j.jcyt.2015.04.008",

language = "English",

volume = "17",

pages = "1104--18",

journal = "Cytotherapy",

issn = "1465-3249",

publisher = "Elsevier",

number = "8",

}

TY - JOUR

T1 - Human adipose-derived stromal cells in a clinically applicable injectable alginate hydrogel

T2 - Phenotypic and immunomodulatory evaluation

AU - Larsen, Bjarke Follin

AU - Juhl, Morten

AU - Cohen, Smadar

AU - Pedersen, Anders Elm

AU - Gad, Monika

AU - Kastrup, Jens

AU - Ekblond, Annette

PY - 2015/8/1

Y1 - 2015/8/1

N2 - BACKGROUND AIMS: Clinical trials have documented beneficial effects of mesenchymal stromal cells from bone marrow and adipose tissue (ASCs) as treatment in patients with ischemic heart disease. However, retention of transplanted cells is poor. One potential way to increase cell retention is to inject the cells in an in situ cross-linked alginate hydrogel.METHODS: ASCs from abdominal human tissue were embedded in alginate hydrogel and alginate hydrogel modified with Arg-Gly-Asp motifs (RGD-alginate) and cultured for 1 week. Cell viability, phenotype, immunogenicity and paracrine activity were determined by confocal microscopy, dendritic cell co-culture, flow cytometry, reverse transcriptase quantitative polymerase chain reaction, Luminex multiplex, and lymphocyte proliferation experiments.RESULTS: ASCs performed equally well in alginate and RGD-alginate. After 1 week of alginate culture, cell viability was >93%. Mesenchymal markers CD90 and CD29 were reduced compared with International Society for Cellular Therapy criteria. Cells sedimented from the alginates during cultivation regained the typical level of these markers, and trilineage differentiation was performed by standard protocols. Hepatocyte growth factor mRNA was increased in ASCs cultivated in alginates compared with monolayer controls. Alginates and alginates containing ASCs did not induce dendritic cell maturation. ASCs in alginate responded like controls to interferon-gamma stimulation (licensing), and alginate culture increased the ability of ASCs to inhibit lymphocyte proliferation.DISCUSSION: ASCs remain viable in alginates; they transiently change phenotype in alginate hydrogel but regain the phenotype of monolayer controls upon release. Cells maintain their paracrine potential while in alginates; the combination of ASCs and alginate is non-immunogenic and, in fact, immunosuppressive.

AB - BACKGROUND AIMS: Clinical trials have documented beneficial effects of mesenchymal stromal cells from bone marrow and adipose tissue (ASCs) as treatment in patients with ischemic heart disease. However, retention of transplanted cells is poor. One potential way to increase cell retention is to inject the cells in an in situ cross-linked alginate hydrogel.METHODS: ASCs from abdominal human tissue were embedded in alginate hydrogel and alginate hydrogel modified with Arg-Gly-Asp motifs (RGD-alginate) and cultured for 1 week. Cell viability, phenotype, immunogenicity and paracrine activity were determined by confocal microscopy, dendritic cell co-culture, flow cytometry, reverse transcriptase quantitative polymerase chain reaction, Luminex multiplex, and lymphocyte proliferation experiments.RESULTS: ASCs performed equally well in alginate and RGD-alginate. After 1 week of alginate culture, cell viability was >93%. Mesenchymal markers CD90 and CD29 were reduced compared with International Society for Cellular Therapy criteria. Cells sedimented from the alginates during cultivation regained the typical level of these markers, and trilineage differentiation was performed by standard protocols. Hepatocyte growth factor mRNA was increased in ASCs cultivated in alginates compared with monolayer controls. Alginates and alginates containing ASCs did not induce dendritic cell maturation. ASCs in alginate responded like controls to interferon-gamma stimulation (licensing), and alginate culture increased the ability of ASCs to inhibit lymphocyte proliferation.DISCUSSION: ASCs remain viable in alginates; they transiently change phenotype in alginate hydrogel but regain the phenotype of monolayer controls upon release. Cells maintain their paracrine potential while in alginates; the combination of ASCs and alginate is non-immunogenic and, in fact, immunosuppressive.

KW - Adipocytes

KW - Adipose Tissue

KW - Adult

KW - Aged

KW - Aged, 80 and over

KW - Alginates

KW - Cell Differentiation

KW - Cell Proliferation

KW - Cell Survival

KW - Cells, Cultured

KW - Coculture Techniques

KW - Female

KW - Glucuronic Acid

KW - Hepatocyte Growth Factor

KW - Hexuronic Acids

KW - Humans

KW - Hydrogel

KW - Immunomodulation

KW - Interferon-gamma

KW - Male

KW - Mesenchymal Stem Cell Transplantation

KW - Mesenchymal Stromal Cells

KW - Middle Aged

KW - Oligopeptides

KW - RNA, Messenger

KW - Tissue Embedding

KW - Young Adult

U2 - 10.1016/j.jcyt.2015.04.008

DO - 10.1016/j.jcyt.2015.04.008

M3 - Journal article

C2 - 26031743

SN - 1465-3249

VL - 17

SP - 1104

EP - 1118

JO - Cytotherapy

JF - Cytotherapy

IS - 8

ER -

Human adipose-derived stromal cells in a clinically applicable injectable alginate hydrogel: Phenotypic and immunomodulatory evaluation

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