Epigenetic Features of Human Perinatal Stem Cells Redefine Their Stemness Potential

Human perinatal stem cells (SCs) can be isolated from fetal annexes without ethical or safety limitations. They are generally considered multipotent; nevertheless, their biological characteristics are still not fully understood. The aim of this study was to investigate the pluripotency potential of human perinatal SCs as compared to human induced pluripotent stem cells (hiPSCs). Despite the low expression of the pluripotent factors NANOG, OCT4, SOX2, and C-KIT in perinatal SC, we observed minor differences in the promoters DNA-methylation profile of these genes with respect to hiPSCs; we also demonstrated that in perinatal SCs miR-145-5p had an inverse trend in comparison to these stemness markers, suggesting that NANOG, OCT4, and SOX2 were regulated at the post-transcriptional level. The reduced expression of stemness markers was also associated with shorter telomere lengths and shift of the oxidative metabolism between hiPSCs and fetal annex-derived cells. Our findings indicate the differentiation ability of perinatal SCs might not be restricted to the mesenchymal lineage due to an epigenetic barrier, but other regulatory mechanisms such as telomere shortening or metabolic changes might impair their differentiation potential and challenge their clinical application.

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The Aberrant DNA Methylation Profile of Human Induced Pluripotent Stem Cells Is Connected to the Reprogramming Process and Is Normalized During In Vitro Culture

PLoS ONE ◽

10.1371/journal.pone.0157974 ◽

2016 ◽

Vol 11 (6) ◽

pp. e0157974 ◽

Cited By ~ 13

Author(s):

Lenka Tesarova ◽

Pavel Simara ◽

Stanislav Stejskal ◽

Irena Koutna

Keyword(s):

Dna Methylation ◽

Stem Cells ◽

In Vitro Culture ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Methylation Profile ◽

Dna Methylation Profile ◽

Aberrant Dna Methylation ◽

Induced Pluripotent

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Mini Review; Differentiation of Human Pluripotent Stem Cells into Oocytes

Current Stem Cell Research & Therapy ◽

10.2174/1574888x15666200116100121 ◽

2020 ◽

Vol 15 (4) ◽

pp. 301-307 ◽

Cited By ~ 3

Author(s):

Gaifang Wang ◽

Maryam Farzaneh

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Embryonic Stem ◽

Human Pluripotent Stem Cells ◽

Human Oocyte ◽

Differentiation Potential ◽

Cell Lineages ◽

Cell Based Therapy ◽

Induced Pluripotent

Primary Ovarian Insufficiency (POI) is one of the main diseases causing female infertility that occurs in about 1% of women between 30-40 years of age. There are few effective methods for the treatment of women with POI. In the past few years, stem cell-based therapy as one of the most highly investigated new therapies has emerged as a promising strategy for the treatment of POI. Human pluripotent stem cells (hPSCs) can self-renew indefinitely and differentiate into any type of cell. Human Embryonic Stem Cells (hESCs) as a type of pluripotent stem cells are the most powerful candidate for the treatment of POI. Human-induced Pluripotent Stem Cells (hiPSCs) are derived from adult somatic cells by the treatment with exogenous defined factors to create an embryonic-like pluripotent state. Both hiPSCs and hESCs can proliferate and give rise to ectodermal, mesodermal, endodermal, and germ cell lineages. After ovarian stimulation, the number of available oocytes is limited and the yield of total oocytes with high quality is low. Therefore, a robust and reproducible in-vitro culture system that supports the differentiation of human oocytes from PSCs is necessary. Very few studies have focused on the derivation of oocyte-like cells from hiPSCs and the details of hPSCs differentiation into oocytes have not been fully investigated. Therefore, in this review, we focus on the differentiation potential of hPSCs into human oocyte-like cells.

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Collagen coated electrospun polyethersulfon nanofibers improved insulin producing cells differentiation potential of human induced pluripotent stem cells

Artificial Cells Nanomedicine and Biotechnology ◽

10.1080/21691401.2018.1508031 ◽

2018 ◽

Vol 46 (sup3) ◽

pp. S734-S739 ◽

Cited By ~ 8

Author(s):

Reyhaneh Nassiri Mansour ◽

Fatemeh Soleimanifar ◽

Mohamad Foad Abazari ◽

Sepehr Torabinejad ◽

Abdolreza Ardeshirylajimi ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Differentiation Potential ◽

Insulin Producing Cells ◽

Induced Pluripotent

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283 GENERATION AND CHARACTERIZATION OF BOVINE INDUCED PLURIPOTENT STEM CELLS

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab283 ◽

2013 ◽

Vol 25 (1) ◽

pp. 289

Author(s):

O. J. Koo ◽

H. S. Kwon ◽

D. K. Kwon ◽

K. S. Kang ◽

B. C. Lee ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Transgenic Animals ◽

Mouse Embryonic Fibroblast ◽

Embryoid Bodies ◽

Differentiation Potential ◽

Serum Replacement ◽

Induced Pluripotent ◽

Large Animals

Stem cells in large animals are an excellent model for cell therapy research and fine resources for producing transgenic animals. However, there are only few reports of stem cells in large animals because of technical differences between species. In this report, we successfully generate bovine induced pluripotent stem cells (iPSC) using 4 human reprogramming factors (Oct4, Sox2, Klf4, and c-myc) under control of PiggyBac transposition vector. Fibroblasts derived from bovine fetuses were transfected using FugeneHD agent. After 21 days, colony-shaped structures on the culture plates were mechanically detached and then seeded on a mouse embryonic fibroblast (MEF) feeder layer pretreated with mitomycin C. The culture medium was DMEM/F12 supplemented with 20% serum replacement, 5 ng mL–1 basic fibroblast growth factor (bFGF), 0.1 mM β-mercaptoethanol, 1% NEAA, and 1% penicillin-streptomycin antibiotics. The iPSC colonies showed alkaline phosphatase activity and expressed several pluripotency markers (Oct4, Sox2, SSEA1, and SSEA4). To confirm differentiation potential, the iPSC were cultured as embryoid bodies and then plated again. βIII-tubulin (ectoderm) and GFAP or α-SMA (mesoderm) were well expressed on the attached cells. The results revealed that the bovine fibroblasts were well inducted to iPSC that had potential of multilineage differentiation. We hope this technology contributes to improving transgenic cattle production. This study was financially supported by IPET (grant # 109023-05-3-CG000, 111078-03-1-CG000) and the BK21 program for Veterinary Science.

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Electrospun silk nanofibers improve differentiation potential of human induced pluripotent stem cells to insulin producing cells

Materials Science and Engineering C ◽

10.1016/j.msec.2019.110398 ◽

2020 ◽

Vol 108 ◽

pp. 110398 ◽

Cited By ~ 2

Author(s):

Seyed Ehsan Enderami ◽

Seyedeh Fatemeh Ahmadi ◽

Reyhaneh Nassiri Mansour ◽

Saeid Abediankenari ◽

Hossein Ranjbaran ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Differentiation Potential ◽

Insulin Producing Cells ◽

Induced Pluripotent

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Long-Term Stability and Differentiation Potential of Cryopreserved cGMP-Compliant Human Induced Pluripotent Stem Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21010108 ◽

2019 ◽

Vol 21 (1) ◽

pp. 108 ◽

Cited By ~ 4

Author(s):

Mehdi Shafa ◽

Tylor Walsh ◽

Krishna Morgan Panchalingam ◽

Thomas Richardson ◽

Laura Menendez ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Genomic Stability ◽

Directed Differentiation ◽

Differentiation Potential ◽

Long Term Stability ◽

Induced Pluripotent ◽

Cell Banks ◽

2D And 3D

The clinical effectiveness of human induced pluripotent stem cells (iPSCs) is highly dependent on a few key quality characteristics including the generation of high quality cell bank, long-term genomic stability, post-thaw viability, plating efficiency, retention of pluripotency, directed differentiation, purity, potency, and sterility. We have already reported the establishment of iPSC master cell banks (MCBs) and working cell banks (WCBs) under current good manufacturing procedure (cGMP)-compliant conditions. In this study, we assessed the cellular and genomic stability of the iPSC lines generated and cryopreserved five years ago under cGMP-compliant conditions. iPSC lines were thawed, characterized, and directly differentiated into cells from three germ layers including cardiomyocytes (CMs), neural stem cells (NSCs), and definitive endoderm (DE). The cells were also expanded in 2D and 3D spinner flasks to evaluate their long-term expansion potential in matrix-dependent and feeder-free culture environment. All three lines successfully thawed and attached to the L7TM matrix, and formed typical iPSC colonies that expressed pluripotency markers over 15 passages. iPSCs maintained their differentiation potential as demonstrated with spontaneous and directed differentiation to the three germ layers and corresponding expression of specific markers, respectfully. Furthermore, post-thaw cells showed normal karyotype, negative mycoplasma, and sterility testing. These cells maintained both their 2D and 3D proliferation potential after five years of cryopreservation without acquiring karyotype abnormality, loss of pluripotency, and telomerase activity. These results illustrate the long-term stability of cGMP iPSC lines, which is an important step in establishing a reliable, long-term source of starting materials for clinical and commercial manufacturing of iPSC-derived cell therapy products.

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Human Endometrial Cells Express Elevated Levels of Pluripotent Factors and Are More Amenable to Reprogramming into Induced Pluripotent Stem Cells

Endocrinology ◽

10.1210/en.2010-1072 ◽

2011 ◽

Vol 152 (3) ◽

pp. 1080-1089 ◽

Cited By ~ 30

Author(s):

Joo Hyun Park ◽

Laurence Daheron ◽

Sibel Kantarci ◽

Byung Seok Lee ◽

Jose M. Teixeira

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Human Endometrium ◽

Skin Fibroblasts ◽

Endometrial Cells ◽

Endogenous Expression ◽

Pluripotent Factors ◽

Neonatal Skin ◽

Induced Pluripotent

The human endometrium is a tissue with remarkable plasticity and regenerative capacity. Additionally, endometrial cells can be retrieved using minimally invasive procedures, which makes them an ideal source for reprogramming into a pluripotent state. Endometrial cells were obtained from donors in their fifth decade and reprogrammed into induced pluripotent stem (iPS) cells using retroviral transduction with SOX2, OCT4, KLF4, and MYC. The human endometrial cells displayed accelerated expression of endogenous NANOG and OCT4 during reprogramming compared with neonatal skin fibroblasts. As a result, iPS cell colonies that could be subcultured and propagated were established as early as 12 d after transduction rather than the usually reported 3–4 wk for other cell types. After 3 wk of reprogramming, the human endometrial cells also yielded significantly higher numbers of iPS colonies in comparison with the neonatal skin fibroblasts. Although the efficiency of iPS colony formation varied depending on the donor, the basal level of endogenous expression of the defined factors was positively correlated with reprogramming efficiency. The reprogramming resulted in an average colony-forming efficiency of 0.49 ± 0.10%, with a range from 0.31–0.66%, compared with the neonatal skin fibroblasts, resulting in an average efficiency of 0.03 ± 0.00% per transduction, with a range from 0.02–0.03%. Our studies show that the human endometrium expresses elevated levels of pluripotent factors, which with additional defined factors, results in significantly more efficient and accelerated generation of induced pluripotent stem cells compared with conventional somatic cells.

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Hematopoietic Differentiation of Human Induced Pluripotent Stem Cells

Blood ◽

10.1182/blood.v112.11.731.731 ◽

2008 ◽

Vol 112 (11) ◽

pp. 731-731

Author(s):

Kyung-Dal Choi ◽

Junying Yu ◽

Kimberly Smuga-Otto ◽

Jessica Dias ◽

Giorgia Salvagiotto ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Differential Expression ◽

Pluripotent Stem Cells ◽

Blood Cells ◽

Hematopoietic Cells ◽

Patient Specific ◽

Hematopoietic Differentiation ◽

Differentiation Potential ◽

Induced Pluripotent

Abstract Induced pluripotent stem cells (iPSCs) provide an unprecedented opportunity for modeling of human diseases in vitro as well as for developing novel approaches for regenerative therapy based on immunologically compatible cells. In the present study, we employed an OP9 differentiation system to characterize the hematopoietic differentiation potential of seven human iPSC lines obtained from human fetal, neonatal, and adult fibroblasts through reprogramming with POU5F1, SOX2, NANOG, and LIN28 and compared it with the differentiation potential of five human embryonic stem cell lines (hESC; H1, H7, H9, H13, and H14). Similar to hESCs, all iPSCs in coculture with OP9 generated all types of colony forming cells (CFCs) as well as CD34+ cells that can be separated into distinct subsets based on differential expression of CD43 and CD31. CD34+CD31+CD43− cells obtained from all iPSCs expressed molecules present on endothelial cells and readily formed a monolayer when placed in endothelial conditions, while hematopoietic CFC potential was restricted to CD43+ cells. iPSC-derived CD43+ cells could be separated into three major subsets based on differential expression of CD235a/CD41a and CD45: CD235a+CD41a+/− (erythro-megakaryocytic progenitors), and lin-CD34+CD43+CD45− (multipotent), and lin-CD34+CD43+CD45+ (myeloid-skewed) primitive hematopoietic cells. Both subsets of primitive hematopoietic cells expressed genes associated with myeloid and lymphoid development, although myeloid genes were upregulated in CD45+ cells, which are skewed toward myeloid differentiation. Cytogenetic analysis demonstrated that iPSCs and derived from them CD43+ cells maintained normal karyotype. In addition short tandem repeat analysis of CFCs generated from IMR90-1 cells has been performed to confirm that blood cells are in fact derived from reprogrammed IMR90 cells, and not from contaminating hESCs. While we observed some variations in the efficiency of hematopoietic differentiation between different iPSCs, the pattern of differentiation was very similar in all seven tested iPSC and five hESC lines. Using different cytokine combinations and culture conditions we were able to expand iPSC-derived myeloid progenitors and induce their differentiation toward red blood cells, neutrophils, eosinophils, macrophages, ostoeclasts, dendritic and Langerhans cells. Although several issues remain to be resolved before iPSC-derived blood cells can be administered to humans for therapeutic purposes, patient-specific iPSCs can already be used for characterization of mechanisms of blood diseases and to identify molecules that can correct affected genetic networks.

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