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Table 2 A methodological pattern of studies on assessing the effects of cell-based products’ implantation into a defect of the vocal folds

From: Stem cell therapy for vocal fold regeneration after scarring: a review of experimental approaches

 

In vitro characterization

Cell type and detection method after IMP

Markers for MA

Histological staining

Markers for IHC

Morphometric parameters

In vivo assessment

Physical properties

Mechanics

Vibration

Kanemaru et al. [49]

FACS (Sca-l, CD29, CD34, CD44, CD45, CD4ge, and 7-AAD)

Xeno (murine) BM-MSC

GFP tracing

No

H&E

Keratin

Desmin

No

No

No

No

Hertegard, et al. [23]

FACS (CD73, CD90, CD105, CD166, CD14, CD31, CD34, CD45, and CD80)

Xeno (human) BM-MSC

FISH

No

H&E

Collagen type I

Collagen density

No

No

No

Lee et al. [45]

No

Auto AD-MSC

CM-Dil tracking

No

H&E

No

No

Direct laryngoscopy

No

No

Johnson et al. 2010 [52]

– FACS (CD11b, CD19, CD-45, 7-AAD)

– AlamarBlue assay

– ICC (CD-44, Sca-1)

Xeno (murine) BM-MSC

GFP tracing

COL3A1

FN1

HAS3

HYAL3

SMA

TGF- β1

No

No

No

No

No

No

Xu et al. [46]

– Phase contrast microscopy

– MTT assay

– FACS (CD34, CD44, CD105, CD106)

– ICC (Vimentin)

– HS (Oil red O, Von Kossa)

– SEM

Auto AD-MSC

DAPI tracking

No

H&E

MT

AB

FN

Collagen, HA, FN density

Endoscopy

No

No

Ohno et al. [50]

No

Auto BM-MSC

ND

No

vG

AB

No

LP thickness

Collagen and HA density

No

No

No

Kim et al. [60]

No

Xeno (murine) BM-MSC

GFP tracing

COL1A1

HAS2

FN1

TGF- β1

H&E

MT

AB

Collagen type I

Collagen and GAG density

Endoscopy

No

No

Kim et al. [38]

– FACS (CD73, CD90, CD31, CD34, CD-45)

– Differentiation test (AG, OG, CG)

Xeno (human) AD-MSC

CM-DiI tracking

No

H&E

MT

VvG

HGF

FSP-1

Collagen type I

FN

Collagen, elastin, and FN density

Endoscopy

No

No

Peng et al. [27]

– FACS (CD29, CD44, CD90, CD105e, CD34, CD45)

– Clonogenicity assay

– Differentiation test (AG, OG, CG)

Allo EM-MSC

Dil tracking

IHC (SMA, vimentin)

No

H&E

MT

vG

AB

FN

Relative content of collagen, elastin and HA

Comparativeendoscopy

No

No

Choi et al. [25]

No

Xeno BM-MSC

In vivo DiD tracking

H&E

MT

AB

PR

 

Scar index measurement

Intensity of collagen optical density

Endoscopy

No

No

Hu et al. [39]

– Phase contrast microscopy

– ICC (vimentin, collagen)

Auto AD-MSC/VF fibroblasts/dAD-MSC

ND

No

No

Collagen

Elastin

HA

Decorin

FN

No

Endoscopy

No

No

Hiwatashi et al. [40]

No

Allo (murine) BM-MSC and AD-MSC

GFP tracing

FGF2 HGF COL1A1

COL3A1

HAS1

HAS2

HAS2

MMP1

VEGFa

vG

AB

No

LP thickness collagen and HA density

No

No

No

Svensson et al. [57]

No

Xeno (human) ESC

FISH

No

H&E

Collagen type I

LP thickness

Collagen density

Scored fibrosis analysis

No

No

No

Valerie et al. [53]

No

Allo AD-MSC

ND

No

H&E

MT

AB

RO

No

LP thickness

Scored collagen, elastin and HA density

No

No

No

De Bonnecaze et al. [41]

– FACS (CD34, CD45, CD31, CD73, CD90)

– Differentiation test (AG, OG)

Auto AD-MSC

GFP tracing via LV transduction

No

H&E

PR

O

Collagen type I

FN

LP and EP thickness

PR optical density

Density of elastic fibers

Number of inflammation foci (foci/mm3)

No

No

No

Shiba et al. [47]

– HA (H&E, vG, MT)

– IHC (vimentin, panCK)

Auto AD-MSC in fibrin gel

ND

No

H&E

vG

MT

No

No

Endoscopy

No

No

Imaizumi, et al. [26]

No

Xeno (human) iPSC

FISH

No

H&E

vG

MT

panCK

AFP

ALPP

Collagen and elastin density

Retention of hydrogel

No

No

No

Morisaki et al. [61]

FACS (CD29, CD90, CD45)

Allo AD-MSC

GFP tracing

HAS1

HAS2

HAS3

COL1A1

COL3A1

MMP1

MMP8

FGF2

HGF

VEGFa

vG

AB

No

Collagen and HA density

No

No

No

Bartlett et al. [54]

No

Xeno (human) BM-MSC

Human β-actin expression

COL1A2

COL3A1

FN1

FMOD

LPL

OCN

HAS

HYAL2

IL1β

IL17β, TGF-β1, TNF

INγ

αSMA

MT

VvG

AB

No

Relative collagen, elastin and HA content

No

No

No

Goel et al. [48]

No

Allo AD-MSC

FISH, PCR (Y CS)

TUNEL labeling for apoptotic cells

No

H&E

MT

No

No

No

No

No

Svistushkin et al. [62]

No

Auto BM-MSC

ND

No

H&E

PR

O

Collagen type I and III

LP thickness

Thickness of collagen fibrils

Scored analysis of 11 morphological parameters

Collagen type I/III ratio

No

No

No

Hertegård et al. [22]

– FACS (CD73, CD90, CD105, HLA-I, CD14, CD34, CD45, HLA-II)

– Migration and invasion assay

– qPCR (HAS, HYAL, MMP)

– ELISA (IL8, HGF, IL6, VEGF, TGF-β1)

Xeno (human) MSC

FISH

No

AB

Collagen type I

Fibrosis, inflammation, MPS intensity

Scored fibrosis analysis

No

No

No

  1. AB Alcian blue, AD-MSC adipose tissue-derived mesenchymal stromal cells, AFP alpha 1 fetoprotein, AG adipogenic, Allo allogeneic, ALPP placental alkaline phosphatase, Auto autological, BM-MSC bone marrow-derived mesenchymal stromal cells, CG chondrogenic, COL1A1 procollagen I type, COL1A2 procollagen alpha 2, COL3A1 procollagen III type, CS chromosome, dAD-MSC differentiated adipose tissue-derived mesenchymal stromal cells, EM epiglottis mucosa, EP epithelium, ESC embryonic stem cells, FGF2 fibroblast growth factor 2, FN1 fibronectin (MA), FMOD fibromodulin, FN fibronectin, FSP-1 fibroblast specific protein 1, GAG glycosaminoglycans, HA hyaluronic acid, HAS1 hyaluronan synthase-I, HAS2 hyaluronan synthase-II, HAS3 hyaluronan synthase-III, HGF hepatocyte growth factor, HLA human leucocyte antigen, HS histological staining, HYAL3 hyaluronidase 3, FACS fluorescence-activated cell sorter analysis, ICC immunocytochemical staining, IHC immunohistochemical staining, IL1β interleukin 1β, IL17β interleukin 17β, INγ interferon γ, iPSC induced pluripotent stem cells, LP lamina propria, LPL lipoprotein lipase, LV lentivirus, MA molecular analysis, MMP1 matrix metalloproteinase 1, MMP8 matrix metalloproteinase 8, MPS mucopolysaccharides, MT Masson trichrome, MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, O orcein, OC osteocalcin, OG osteogenic, panCK pancytokeratin, PR picrosirius red, RO reticulin and orcein, SEM scanning electron microscopy, SMA smooth muscle actin, TGF-β1 transforming growth factor-beta 1, TNF tumor necrosis factor, VEGFa vascular endothelial growth factor type A, vG van Gieson, VvG Verhoeff-Van Gieson, Xeno xenogeneic