Journal of Stem Cells & Regenerative Medicine最新文献

Human pluripotent stem cells (hPSCs) are a promising source of somatic cells for clinical applications and disease modelling. However, during culture they accumulate genetic aberrations such as amplification of 20q11.21 which occurs in approximately 20% of extensively cultured hPSC lines and confers a BCL2L1-mediated survival advantage. During the production of the large number of cells required for transplantation and therapy these aberrations may become unavoidable which has important safety implications for therapies and may also impact upon disease modelling. Presently, these risks are poorly understood; whilst it is apparent that large-scale genetic aberrations can pose an oncogenic risk, the risks associated with smaller, more insidious changes have not been fully explored. In this report, the effects of engraftment of human embryonic stem cells (hESCs) and hESC-derived hepatocyte-like cells (HLCs) with and without amplification of the 20q11.21 minimal amplicon and isochromosome 20q (i20q) in SCID-beige mice are presented. The cells were tracked in vivo using a luminescent reporter over a period of approximately four months. Intrasplenic injection of hESCs showed greater engraftment potential and the formation of more severely disruptive lesions in the liver and spleen of animals injected with cells containing 20q11.21 compared with i20q and wild type. HLCs with 20q11.21 engrafted more successfully and formed more severely disruptive lesions than wild type cells or cells with i20q. These results reinforce the notion that karyotyping of therapeutic hPSC is required for transplant, and suggest that screening for known common aberrations is necessary. Further work to identify commonly arising genetic aberrations should be performed and routine screening for hPSCs intended for therapeutic use should be used.

The goals of treatment for fingertip injuries are maximising digital length, tactile sensation, pulp padding, and fingertip appearance while minimising complications like infection and amputation. Currently, terminalisation, healing by secondary intention, and flap surgeries are widely used for crushing fingertip injuries, but they have their own set of issues and limitations. We present a tissue-engineered method by combining platelet-rich fibrin injections with stacked-up layers of synthetic biodegradable temporising matrix to treat a severely crushed fingertip. This novel therapy minimised reconstructions while successfully regenerating new soft-tissues. Soft-tissue regeneration within the stacked-up biodegradable matrix achieved adequate volume, sensation, function, and mobility of the newly reconstructed fingertip while maintaining its skeletal length. Notably, the regenerated fingertip allowed the patient to resume work normally as a busy software engineer. Thus, minimalistic fingertip reconstruction not only prevented a disability, but also served as a viable alternative to major reconstructive surgeries.

Coronary heart disease (CHD) is a leading cause of death globally, while its current management is limited to reducing the myocardial infarction area without actually replacing dead cardiomyocytes. Direct cell reprogramming is a method of cellular cardiomyoplasty which aims for myocardial tissue regeneration, and CD34⁺ cells are one of the potential sources due to their shared embryonic origin with cardiomyocytes. However, the isolation and culture of non-adherent CD34⁺ cells is crucial to obtain adequate cells for high-efficiency genetic modification. This study aimed to investigate the optimal method for isolation and culture of CD34⁺ peripheral blood cells using certain culture media. A peripheral blood sample was obtained from a healthy subject and underwent pre-enrichment, isolation, and expansion. The culture was subsequently observed for their viability, adherence, and confluence. Day 0 observation of the culture showed a healthy CD34⁺ cell with a round cell shape, without any adherent cells present yet. Day 4 of observation showed that CD34⁺ cells within the blood plasma medium became adherent, indicated by their transformations into spindle or oval morphologies. Meanwhile, CD34⁺ cells in vitronectin and fibronectin media showed no adherent cells and many of them died. Day 7 observation revealed more adherent CD34⁺ cells in blood plasma medium, and which had 75% of confluence. In conclusion, the CD34⁺ cells that were isolated using a combination of density and magnetic methods may be viable and adequately adhere in culture using blood plasma medium, but not in cultures using fibronectin and vitronectin.

Mesenchymal stem cells derived from adipose tissue (ADMSCs) are being increasingly considered in regenerative medicine-based clinical applications. Apart from possessing therapeutic applications themselves, ADMSCs also secrete a myriad of soluble factors which are promising candidates for treating several degenerative diseases such as osteoarthritis and neurodegenerative diseases, wound repair as well as for cosmeceutical purposes. In our research study, we successfully isolated ADMSCs in-house, now called CKC-Endeavour-1 from the lipoaspirate sample of a patient who underwent liposuction. The subsequent expansion of cells was performed in xeno-free and serum-free conditions and their characterisation was performed using tri-lineage differentiation studies. The levels of differentiation were assessed by staining and gene expression which was observed to be comparable between the in-house developed ADMSC cell line and the commercially purchased ADMSCs. Following characterisation, the secretory components from these MSCs, namely, conditioned media (ADMSC-CM) and exosomes (ADMSC-EXO) were harvested from CKC-Endeavour-1 under xeno-free, serum-free, and supplement-free conditions followed by lyophilisation in order to attempt to prolong its shelf-life. The comprehensive analysis of the secretome profile of ADMSC-CM using carried out using cytokine array and demonstrated the presence of 105 cytokines and growth factors. Also, clinical grade Izon columns were used to isolate the exosomes from ADMSC-CM obtaining exosomes in the size range of <200nm, analysed using nanoparticle tracking analysis. Overall, our study developed an ADMSC cell line, CKC-Endeavour-1, along with their CM and exosome (EXO) products under clinically safe conditions. Additionally, we have obtained a comprehensive understanding of the secreted factors present in the ADMSC-CM which could be further explored in detail to tap the best therapeutic benefits from them.

Orthobiologics never cease to cause popularity within the medical science field, distinctly in regenerative medicine. Recently, adipose tissue has been an object of interest for many researchers and medical experts due to the fact that it represents a novel and potential cell source for tissue engineering and regenerative medicine purposes. Stromal vascular fraction (SVF), for instance, which is an adipose tissue-derivative, has generated optimistic results in many scenarios. Its biological potential can be harnessed and administered into injured tissues, particularly areas in which standard healing is disrupted. This is a typical feature of osteoarthritis (OA), a common degenerative joint disease which is outlined by persistent inflammation and destruction of surrounding tissues. SVF is known to carry a large amount of stem and progenitor cells, which are able to perform self-renewal, differentiation, and proliferation. Furthermore, they also secrete several cytokines and several growth factors, effectively sustaining immune modulatory effects and halting the escalated pro-inflammatory status of OA. Although SVF has shown interesting results throughout the medical community, additional research is still highly desirable in order to further elucidate its potential regarding musculoskeletal disorders, especially OA.

Background: Mesenchymal stem cells are currently used to treat several diseases. Populations of putative stem cells found in the adipose tissue (ASCs) have been shown to possess particularly enhanced functionalities. Nonetheless, there is lack of evidence that evaluates the effects of cryopreservation techniques on well-defined functional ASC populations characterized by immunophenotypical repertoire.

Objective: We therefore embarked a study to compare the frozen-thawed ASC subsets: CD73+CD90+CD105+CD34+CD146-(CD34+CD146), CD73+CD90+CD105+CD34+CD146+(CD34+CD146+), and CD73+CD90+CD105+CD34+(CD34+). We assessed their characterization in different functional assays.

Method: The ASC immunophenotypical subsets-purified by a flow cytometry sorting technique-were frozen in liquid nitrogen. After a period, they were thawed to examine their differentiation ability, colony-forming units, viability, and growth rate.

Results: We confirmed that inside the primary cell culture system, the proportion of CD34+, CD34+CD146-, and CD34+CD146+ took up 80%, 62%, and 19% on average, respectively. All populations could be frozen and stored in liquid nitrogen with retention of more than 85% of cell viability and displayed comparable stemness characteristics. Most importantly, the CD34+CD146+ subpopulation displayed a higher proliferation rate than other groups.

Conclusion: Our data demonstrated that the frozen-thawed CD34+CD146+ cells might represent a promising source for autologous cellular-based therapy. These findings set the basis for ASC subpopulations-based application in future potential clinical settings.