Submit a Manuscript: World J Stem Cells 2019 August 26; 11(8): 506-518 DOI: 10.4252/wjsc.v11.i8.506ISSN 1948-0210 (online)
REVIEW Tonsil-derived stem cells as a new source of adult stem cells
Kyung-Ah Cho, Hyun Jung Lee, Hansaem Jeong, Miri Kim, Soo Yeon Jung, Hae Sang Park, Kyung-Ha Ryu, Seung Jin Lee, Byeongmoon Jeong, Hyukjin Lee, Han Su Kim
ORCID number: Kyung-Ah Cho (0000-0003-3758-4209); Hyun Jung Lee (0000-0002-3573-0308); Hansaem Jeong (0000-0001-7081-2088); Miri Kim (0000-0002-7078-4168); Soo Yeon Jung (0000-0001-7497-3057); Hae Sang Park
(0000-0002-5968-2507); Kyung-Ha Ryu (0000-0001-8424-2303); Seung Jin Lee (0000-0003-1216-0688); Byeongmoon Jeong
(0000-0001-9582-1343); Hyukjin Lee (0000-0001-9478-8473); Han Su Kim (0000-0003-2239-0225).
Author contributions: Cho KA and Lee HJ contributed equally to writing this paper as the first authors; Jeong H, Kim M, Jung SY and Park HS contributed to the literature review, analysis, and artwork.; Ryu KH and Lee S contributed to the conception and design of the study. Jeong B, Lee H, and Kim HS equally contributed to the drafting, revision, and editing of the manuscript, and gave approval to the final version as corresponding authors. Supported by the Korea Health Technology RD Project through the Korea Health Industry Development Institute; the Ministry of Health and Welfare, No. HI16C- 2207; the Basic Science Rearch Program through the NRF, No. NRF-
2018R1D1A1A09083264; Ewha Womans University, No. RP-grant 2017.
Conflict-of-interest statement: There are no potential conflicts of interest to report.
Open-Access: This is an open-access article that was lected by an in-hou editor and fully peer-Kyung-Ah Cho, Department of Microbiology, College of Medicine, Ewha Womans University, Seoul 07985, South Korea
Hyun Jung Lee, Byeongmoon Jeong, Department of Chemistry and Nano Science, Ewha Womans University, Seoul 03760, South Korea
Hansaem Jeong, Miri Kim, Seung Jin Lee, Hyukjin Lee, College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul 03760, South Korea
Soo Yeon Jung, Han Su Kim, Department of Otorhinolaryngology, College of Medicine, Ewha Womans University, Seoul 07985, South Korea
Hae Sang Park, Department of Otorhinolaryngology, College of Medicine, Hallym University, Chuncheon 24252, South Korea
Kyung-Ha Ryu, Department of Pediatrics, College of Medicine, Ewha Womans University, Seoul 07985, South Korea态度决定一切英文
Corresponding author: Han Su Kim, MD, PhD, Professor, Department of Otorhinolaryngology, College of Medicine, Ewha Womans University, Ahnyangcheon-ro 1071 Seoul 07985, South Korea. vent@ewha.ac.kr
Telephone: +82-10-87185316路由器安装教程
Fax: +82-2-26535135
Abstract
Located near the oropharynx, the tonsils are the primary mucosal immune organ. Tonsil tissue is a promising alternative source for the high-yield isolation of adult stem cells, and recent studies have reported the identification and isolation of tonsil-derived stem cells (T-SCs) from waste surgical tissue following tonsillectomies in relatively young donors (i.e., under 10 years old). As such, T-SCs offer veral advantages, including superior proliferation and a shorter doubling time compared to bone marrow-derived menchymal stem cells (MSCs). T-SCs also exhibit multi-lineage differentiation, including mesodermal, endodermal (e.g., hepatocytes and parathyroid-like cells), and even ectodermal cells (e.g., Schwann cells). To this end, numbers of rearchers have evaluated the practical u of T-SCs as an alternative source of autologous or allogenic MSCs. In this review, we summarize the details of T-SC isolation and identification and provide an overview of their application in cell therapy and regenerative medicine.
Key words: Stem cell; Tonsil-derived stem cell; Differentiation; Endoderm; Mesoderm; Ectoderm; Cell therapy
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s/by-nc/4.0/Manuscript source : Invited
Manuscript
Received: February 13, 2019
Peer-review started: February 15,
2019First decision: March 26, 2019
Revid: May 31, 2019
Accepted: July 29, 2019
Article in press: July 29, 2019
Published online: August 26, 2019
P-Reviewer: Liu L, Li SC, Saeki K,
Tanabe S, Wakao H
S-Editor: Cui LJ
L-Editor: A
E-Editor: Xing YX ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights rerved.Core tip: The u of adult stem cells is often limited by the lack of differentiation among stem cells isolated from certain germ layers. However, tonsil-derived stem cells (T-SCs)were able to differentiate into various tissue types from the three germ layers, which is the most advantageous fe
ature of this new stem cell source. T-SCs can also be ud as native cells in the treatment of various immune-related dias. As a result, it can be concluded that T-SCs have great potential for clinical applications in cell therapy and regenerative medicine.
Citation: Cho KA, Lee HJ, Jeong H, Kim M, Jung SY, Park HS, Ryu KH, Lee SJ, Jeong B,Lee H, Kim HS. Tonsil-derived stem cells as a new source of adult stem cells. World J Stem Cells 2019; 11(8): 506-518URL : /1948-0210/full/v11/i8/506.htm DOI : /10.4252/wjsc.v11.i8.506
INTRODUCTION Recent achievements in the identification, isolation, in vitro culture, and differenti-ation of various adult stem cells are indicative of the unprecedented potential of the cells in treating various degenerative dias [1]. Menchymal stem cells (MSCs) in particular, have been ud clinically for more than 10 years. From animal studies to
clinical trials, MSCs have demonstrated great promi in treating numerous dias,
particularly tissue injury and immune disorders [2]. To obtain the large volumes of cells
required for testing and treatment, various tissue sources have been investigated for
the isolation of MSCs, including bone marrow, adipo tissue, umbilical cord blood,
amniotic fluid, the placenta, dental pulp, and urine [3]. However, the isolation yields of制药工程专业就业方向
MSCs from different tissue sources vary greatly, and the differentiation potential,
yield, and maximal lifespan of isolated MSCs decrea significantly with donor age.
Therefore, it is important to locate new adult stem cell sources to overcome the
limitations.
The human tonsils are located near the oropharynx (palatine tonsils) and
nasopharynx (adenoid), which are part of the respiratory and digestive system. Tonsil
tissue is one of the primary nsitization systems for the generation of B cells, and
tonsil tissue is easily obtained from tonsillectomies, a minimally invasive surgery
conducted most often on patients aged between 5 and 19. Tonsil-derived stem cells (T-
SCs) were first introduced by Janjanin et al [4]. Due to the younger donors, the isolation
yields of T-SCs are much higher than tho from other tissue types. Therefore, T-SCs
have received much attention as alternative allogeneic or autologous cell sources for
clinical u. In this review, we highlight recent rearch on the isolation and
development of T-SCs, which provides strong evidence of their superior
characteristics. In addition to their high proliferation and expansion capacity, T-SCs
can undergo differentiation into cells from all three germ layers (i.e ., ectoderm,
mesoderm, and endoderm). This unique differentiation potential is described in
detail. Finally, we provide an in-depth discussion of the u of T-SCs in cell therapy and regenerative medicine.女短发型
ISOLATING AND IDENTIFYING TONSIL-DERIVED MSCS
Isolating T-SCs consists of two major steps: Enzymatic disaggregation and density
gradient centrifugation [5]. Briefly, small pieces of tonsillar tissues were expod to
enzymes, including collagena type I and DNa for 30 min at 37 °C under stirring.
This solution was then filtered through a wire mesh and 70-µm cell strainer to collect
single-cell suspensions. The mononuclear cell (MNC) fraction was obtained using
Ficoll-Paque (GE Healthcare, Little Chalfont, United Kingdom) density gradient
centrifugation. The MNCs were plated at the density of 108 cells in a T-150 culture
flask with Dulbecco’s modified Eagle’s medium-high gluco (DMEM-HG;
Invitrogen) supplemented with fetal bovine rum and antibiotics. The primary
culture (passage 0; P0) was cultivated until the adherent cells reached confluence and
were passaged by trypsinization (Trypsin, Life Technologies GmbH, Vienna, Austria).
Cho KA et al. Tonsil stem cells
Cho KA et al. Tonsil stem cells
Immunophenotype characterization, which is bad on the expression of cell
surface markers, is the most common method for distinguishing different cell clusters.
To date, extensive rearch has identified various cell surface markers that
characterize the MSCs derived from different sources. In order to identify T-SCs as a
cellular source for new adult MSCs, T-SC surface makers were investigated[5,6]. As
with other MSCs, T-SCs expresd the standard positive markers for MSCs, CD73,
CD90, CD105, CD29, CD44, CD166, CD58, and CD49e. Most of the markers
reprent cellular adhesion molecules which possibly render MSCs to act on other cell
types via direct interaction. On the other hand, T-SCs were negative for the
hematopoietic markers CD14, CD34, CD45, and CD133, the endothelial marker CD31,
and co-stimulatory proteins such as the antigens CD40, CD80, and CD86. In addition,
class II MHC antigens are entirely abnt on T-SCs[5,6]. Becau tonsil tissue is part of
the mucosal immune system and contains large numbers of follicular dendritic cells
(FDCs), additional rearch has been carried out to verify the lack of FDC markers
CD11b, CD21, CD23, CD35, and CD54 in T-SCs[5,7] to confirm no-contamination with
FDC. FDCs are known to originate from tonsillar stromata and proliferate on and
adhere to plastic in vitro. Therefore, the lack of the markers is an important indicator
that can be ud to distinguish T-SCs from FDCs.
MAJOR ADVANTAGES OF T-SCS OVER BONE MARROW-
DERIVED STEM CELLS
Although MSCs can be isolated from various tissue types, they were initially
harvested from bone marrow (BM), which requires a highly invasive procedure[4].
Here, we highlight the significant benefits of using T-SCs in terms of isolation and
clinical u compared with BM-MSCs.
Isolating BM-MSCs has veral limitations, including donor morbidity, and they
are challenging to harvest, thus requiring a high degree of skill. Bone marrow
extraction takes approximately two hours under general anesthesia and requires the
hospitalization and recovery of the donor. Therefore, it is always difficult to find a
sufficient number of donors. In contrast, T-SCs are easily obtained from discarded
tissue; more than 530000 tonsillectomies are performed annually in children younger
than 15 years in the United States[8], meaning that tonsils are one of the most abundant
tissue sources for stem cell isolation.
The age of the donor affects the isolation yield of MSCs, with the number of MSCs
harvested from bone marrow decreasing with donor age. For example, infants have
one colony forming units-fibroblast (CFU-f) per 10000 cells in bone marrow, but this
falls to 1 per 400000 in donors in their 50 s[9]. In contrast, approximately 8-10 × 108
MSCs are isolated from one-third of one tonsil (2 cm x 1.5 cm x 1.5 cm) from donors
under 10 years old[5].
When compared with BM-MSCs, T-SCs offer superior stem cell properties, such as
high lf-renewal and proliferation. For example, T-SCs show a doubling time of 37.1
± 3.4 h for an initial population, compared to 58.2 ± 2.3 h for BM-MSCs[4]. Other
rearch has also confirmed the more rapid proliferation of T-SCs compared with
MSCs derived from adipo tissue[10].
The proliferation of BM-MSCs gradually decreas with passage number, whereas
T-SCs retain their physiological properties for much longer. In general, most cells
become more prominent, longer, less defined, and less proliferative during long-term
in vitro culture as they experience nescence. T-SCs also exhibit the signs of
nescence from passage 7, but the cells proliferate up to passage 15 with no change in
the MSC markers. Tonsil tissue contains as many B cells and T cells as immune
organs, and the cells affect the immune modulation of stem cells. Pro-inflammatory
海上日出课文原文cytokines may also affect the positive differentiation and proliferation of T-SCs[4,11,12],
and this has been supported by rearch on tissue obtained from tonsillectomies in
respon to chronic bacterial infections and chronic tonsillitis[13-15].
Bone marrow and adipo tissue originate from the mesoderm layer, whereas tonsil
tissue has two origins: The epithelial cells derive from the cond pharyngeal pouch
in the endoderm layer, and lymphoid tissue comes from the mesoderm layer, which
invades during fetal development. Rearch has confirmed that T-SCs can be easily
differentiated into endodermal, ectodermal, and mesodermal cells (Figure 1).
THERAPEUTIC POTENTIAL OF T-SCS BASED ON THEIR
DIFFERENTIATION PROPERTIES
Figure 1 Possible reason enables diversity in tonsil-derived stem cells differentiation; constitutive features
of tonsil. Zygote has most powerful differentiation potential (totipotent). It was able to differentiate to all human
anatomies and to become human body. Embryonic stem cells (ESCs) are derived from the inner cell mass of the
early embryo. ESCs also has great developmental potential and was able to differentiate to all cell lineages of an
organism except for extraembryonic tissues (pluripotent). It is well known that ectodermal or endodermal
differentiation is often difficult to achieve with menchymal stem cells isolated from bone marrow and adipo tissue
(multipotent). Tonsil tissues consist of two different origin tissues; epithelial cells from endoderm origin and lymphoid
tissues from mesoderm origin.
Cell therapy and tissue engineering have been investigated to regenerate lost or
malfunctioning organs. The approaches utilize biomaterial scaffolds and MSCs to
facilitate initial cell adhesion and retention while promoting cell growth for tissue
regeneration [16,17]. In particular, the differentiation properties of MSCs are of great
importance for tissue regeneration. It is generally known that isolated MSCs are often
怎样变高
limited to germ-layer specific differentiation. As mentioned earlier, T-SCs offer
multipotent differentiation potential that can be applied in regenerating various tissue types without concern for their germ layer origin.
ECTODERMAL DIFFERENTIATION OF T-SCS
Ectodermal differentiation is often difficult to achieve with MSCs isolated from bone
marrow and adipo tissue. However, under the right conditions, T-SCs can be客厅电视墙颜色
differentiated into non-menchymal lineages, including ectodermal differentiation
into neurons, astrocytes, and Schwann-like cells to support nerve regeneration.
Neuronal differentiation of T-SCs
The neuronal differentiation of T-SCs was investigated in a three dimensional (3D)
hybrid scaffold system by Patel et al [18]. This scaffold was fabricated by increasing the
temperature of an aqueous solution of poly (ethylene glycol)-poly(L-alanine) to 37 °C,
thus instigating the heat-induced sol-to-gel transition, in which T-SCs and growth
factor-releasing microspheres were suspended. The gel exhibited a modulus of 800 Pa
at 37 °C, similarly to that of brain tissue, and was robust enough to hold the microspheres and cells within the 3D cell culture. Neuronal growth factors were
Cho KA et al. Tonsil stem cells
Figure 2 Differentiation potential of tonsil-derived stem cells. The multi-potency of the tonsil-derived stem cells (T-SCs) is confirmed in many studies. Under the proper conditions and signals (box), T-SCs can be differentiated into non-menchymal lineage, such as ectodermal differentiation (neuron
s, and Schwann-like cells)and endodermal cells (parathyroid like cells, islet cells and hepatocytes). T-MSCs: Tonsil- menchymal stem cells.
轮胎常识relead over 12–18 d, and the encapsulated T-SCs gradually exhibited morphological
changes from spherical to multipolar elongation. Significantly higher expression
levels of neuronal biomarkers such as nuclear receptor-related protein, neuron-
specific enola, microtubule-associated protein-2, neurofilament-M, and glial
fibrillary acidic protein were obrved at both the mRNA and protein level in the
hybrid system. This study clearly demonstrates the advantages of 3D hybrid scaffolds
and highlights the importance of the sustained relea of growth factors from hybrid
systems to support the neuronal differentiation of T-SCs.
Schwann cell differentiation of T-SCs
Schwann cells are the glial cells of peripheral nerves that wrap around the axons to
form myelin in the peripheral nervous system. Schwann cells promote nerve
regeneration by creting trophic support molecules and establishing a supportive
growth matrix [19]. Jung et al [20] demonstrated that T-SCs could be differentiated into
Schwann-like cells over veral steps. Briefly, T-SCs were induced to form
neurospheres under stimulation with EGF, bFGF, and B27 for 7 d. The neurospheres
were then triturated and re-plated onto laminin-coated dishes with Schwann cell
differentiation medium. After 10 d of culturing, the cells exhibited morphological
changes, including the formation of elongated bipolar and tripolar spindle shapes.
Schwann-like cells differentiated from T-SCs highly express the Schwann cell markers
GFAP, NGFR, S100B, KROX20, and KROX24. Notably, Schwann cells differentiated
from T-SCs were able to produce myelinate axons in vitro when co-cultured with
mou dorsal root ganglion neurons. In a mou model with a sciatic nerve injury, a
marked improvement in gait and incread nerve regeneration were obrved with
Schwann-cell treatment. Therefore, T-SCs can be a uful source for Schwann cell-bad cell therapy to treat neuropathic dias.
MESODERMAL DIFFERENTIATION OF T-SCS
Mesodermal differentiation is mainly achieved with adult MSCs from the mesodermal
germ layer. Previously, a variety of cell sources from bone marrow and adipo tissue
was utilized for mesodermal differentiation to treat bone, cartilage, and fat disorders.
In this ction, we highlight the potential u of T-SCs as an alternative cell source for
mesodermal differentiation (Figure 2), and we provide a comparative study that illustrates the advantages of T-SCs.
Cho KA et al. Tonsil stem cells
