Comprehensive characterization of human adipose tissue-derived stem cells expanded in vitro

Biologia ◽  
2013 ◽  
Vol 68 (4) ◽  
Author(s):  
Ľuboš Danišovič ◽  
Marcela Kuniaková ◽  
Zuzana Varchulová-Nováková ◽  
Martin Boháč ◽  
Ivan Varga ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Growth Curve ◽  
Morphological Characteristics ◽  
Human Adipose Tissue ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Tem Analysis ◽  
Hla Dr

AbstractAdipose tissue seems to be a rich and safe source of mesenchymal stem cells (MSCs). The present study was aimed to investigate the biological and morphological characteristics of human adipose tissue-derived stem cells (ATSCs). Light and transmission electron microscopy were used. Course of proliferation was analyzed by growth curve. Expression of surface antigens was assessed by flow cytometry. Chondrogenic potential was assessed by immunohistochemistry. Obtained results showed morphology typical of fibroblastoid cells. TEM analysis proved ultrastructural morphology similar to MSCs from other sources. ATSCs reflected their proteosynthetic and metabolic activity. Each cell had irregular shape of nucleus with noticeable nucleoli. Abundant cisterns of rough endoplasmic reticulum were present in their cytoplasm. Karyotype mapping showed normal count of human chromosomes (46,XX). The growth curve revealed high capability for proliferation and population doubling time was 27.36 hours. ATSCs were positive for CD13, CD29, CD44, CD73, CD90, CD105 and CD106, but did not express CD14, CD34, CD45 and HLA-DR. It was also proved that ATSCs underwent chondrogenic differentiation in vitro. On the basis of obtained results it should be emphasized that ATSCs are typical MSCs and after further investigations they may be used in tissue engineering and regenerative medicine.

Culture Expanded Canine Mesenchymal Stem Cells Possess Osteochondrogenic Potential in Vivo and in Vitro

1997 ◽  
Vol 6 (2) ◽  
pp. 125-134 ◽  
Author(s):  
S. Kadiyala ◽  
R. G. Young ◽  
M. A. Thiede ◽  
S. P. Bruder
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Morphological Characteristics ◽  
Cartilage Regeneration ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Human Mscs ◽  
And Cartilage

Mesenchymal Stem Cells (MSCs) possessing the capacity to differentiate into various cell types such as osteoblasts, chondrocytes, myoblasts, and adipocytes have been previously isolated from the marrow and periosteum of human, murine, lapine, and avian species. This study documents the existence of similar multipotential stem cells in canine marrow. The cells were isolated from marrow aspirates using a modification of techniques previously established for human MSCs (hMSCs), and found to possess similar growth and morphological characteristics, as well as osteochondrogenic potential in vivo and in vitro. On the basis of these results, the multipotential cells that were isolated and culture expanded are considered to be canine MSCs (cMSCs). The occurrence of cMSCs in the marrow was determined to be one per 2.5 × 104 nucleated cells. After enrichment of the cMSCs by centrifugation on a Per-coll cushion, the cells were cultivated in selected lots of serum. Like the hMSCs, cMSCs grew as colonies in primary culture and on replating, grew as a monolayer culture with very uniform spindle morphology. The population doubling time for these cMSCs was approximately 2 days. The morphology and the growth kinetics of the cMSCs were retained following repeated passaging. The osteogenic phenotype could be induced in the cMSC cultures by the addition of a synthetic glucocorticoid, dexamethasone. In these osteogenic cultures, alkaline phosphatase activity was elevated up to 10-fold, and mineralized matrix production was evident. When cMSCs were loaded onto porous ceramics and implanted in autologous canine or athymic murine hosts, copious amounts of bone and cartilage were formed in the pores of the implants. The MSC-mediated osteogenesis obtained by the implantation of the various MSC-loaded matrix combinations is the first evidence of osteogenesis in a canine model by implantation of culture expanded autologous stem cells. The identification and isolation of cMSCs now makes it feasible to pursue preclinical models of bone and cartilage regeneration in canine hosts.

scholarly journals in Vitro Effect of Methamphetamine on Proliferation, Differentiation and Apoptosis of Adipose Tissue Stem Cells

2022 ◽  
Vol 25 ◽  
pp. 69-76
Author(s):  
Leyla Anari ◽  
Davood Mehrabani ◽  
Mahboobeh Nasiri ◽  
Shahrokh Zare ◽  
Iman Jamhiri ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Public Health Importance ◽  
Isolated Cells ◽  
Ki 67 ◽  
Proliferation And Differentiation ◽  
Vitro Effect

Purpose: Among abused substances, methamphetamine is a psychostimulant drug widely used recreationally with public health importance. This study investigated the effect of methamphetamine on proliferation, differentiation, and apoptosis of human adipose tissue stem cells (AdSCs). Methods: AdSCs were isolated from human abdominal adipose tissue and were characterized for mesenchymal properties and growth kinetics. MTT assay was undertaken to assess methamphetamine toxicity on proliferation and differentiation properties and apoptosis of hAdSCs. Results: Isolated cells were shown to have mesenchymal properties and a population doubling time (PDT) of 40.1 h. Following methamphetamine treatment, expressions of KI-67 and TPX2 as proliferation genes and Col1A1 and PPARg as differentiation genes decreased. Methamphetamine administration increased the expression of Bax and decreased Bcl-2 genes responsible for apoptosis. Conclusions: Our data suggested when AdSCs were exposed to methamphetamine, it decreased proliferation and differentiation properties of stem cells together with an increase in apoptosis. These findings can be added to the literature, especially when methamphetamine is used recreationally for weight loss purposes.

Hepatocyte differentiation of mesenchymal stem cells from human adipose tissue in vitro promotes hepatic integration in vivo

Gut ◽  
2008 ◽  
Vol 58 (4) ◽  
pp. 570-581 ◽  
Author(s):  
H Aurich ◽  
M Sgodda ◽  
P Kaltwasser ◽  
M Vetter ◽  
A Weise ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Human Adipose Tissue ◽  
Hepatocyte Differentiation ◽  
Adipose Tissue In Vitro

scholarly journals Low Oxygen Tension Potentiates Proliferation and Stemness but Not Multilineage Differentiation of Caprine Male Germline Stem Cells

2021 ◽  
Author(s):  
Shiva Pratap Singh ◽  
Suresh Dinkar Kharche ◽  
Manisha Pathak ◽  
Ravi Ranjan ◽  
Yogesh Kumar Soni ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ex Vivo ◽  
Growth Characteristics ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Germline Stem Cells ◽  
Multilineage Differentiation ◽  
Low Oxygen ◽  
Differentiation Potential

Abstract The milieu of testicular germline stem cells (mGSCs) is characterized as low oxygen (O2) environment, whereas, there in-vitro expansion is typically performed under normoxia (20-21% O2). Here, we evaluated and compared the culture and multilineage differentiation characteristics of enriched (through differential platting and percoll density centrifugation) caprine mGSCs (cmGSCs) under hypoxic (5% O2) and normoxic (21% O2) culture conditions. For this, in addition to growth characteristics and population-doubling time (PDT); viability, proliferation, senescence, and expression of key-markers of adhesion (β-integrin and E-Cadherin) and stemness (OCT-4, THY-1 and UCHL-1) were evaluated and compared under normoxia and hypoxia. Moreover, the extent of multilineage differentiation (neurogenic, adipogenic, and chondrogenic differentiation) was assessed. The survival, viability and proliferation were significantly promoted and PDT was reduced (p < 0.05), thus yielding a higher number of viable cells with larger colonies under hypoxia. Furthermore, expression of stemness and adhesion markers was distinctly increased under lowered O2 condition. Conversely, the presence of differentiated regions and expression of differentiation specific key genes [C/EBPα (adipogenic), nestin and β-tubulin (neurogenic), and COL2A1 (chondrogenic)] were significantly (p < 0.05) reduced under hypoxic conditions. These data demonstrate that culturing cmGSCs under hypoxia augments the growth characteristics, and stemness but not the multilineage differentiation potential of cmGSCs as compared with normoxia. These data are important for the development of robust methodologies for ex-vivo expansion and lineage-committed differentiation of cmGSCs for clinical applications.

Gelatin/Hydroxyapatite Scaffolds: Studies on Adhesion, Growth and Differentiation of Mesenchymal Stem Cells

2010 ◽  
Vol 93-94 ◽  
pp. 121-124
Author(s):  
Nuttapon Vachiraroj ◽  
Siriporn Damrongsakkul ◽  
Sorada Kanokpanont
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Calcium Content ◽  
Population Doubling ◽  
Population Doubling Time ◽  
3 Dimensional

In this work, we developed a 3-dimensional bone tissue engineering scaffold from type B gelatin and hydroxyapatite. Two types of scaffolds, pure gelatin (pI~5) (Gel) and gelatin/hydroxyapatite (30/70 wt./wt.) (Gel/HA), were prepared from concentrated solutions (5% wt./wt.) using foaming/freeze drying method. The results SEM revealed the interconnected-homogeneous pores of Gel and Gel/HA were 121  119 and 148  83m, respectively. Hydroxyapatite improved mechanical property of the gelatin scaffolds, especially at dry state. Compressive modulus of Gel and Gel/HA scaffolds were at 118±21.68 and 510±109.08 kPa, respectively. The results on in vitro cells culture showed that Gel/HA scaffolds promoted attachment of rat’s mesenchymal stem cells (MSC) to a 1.23 folds higher than the Gel scaffolds. Population doubling time (PDT) of MSC on Gel and Gel/HA scaffolds were 51.16 and 54.89 hours, respectively. In term of osteogenic differentiation, Gel/HA scaffolds tended to enhance ALP activity and calcium content of MSC better than those of the Gel scaffold. Therefore the Gel/HA scaffolds had a potential to be applied in bone tissue engineering.

Paradoxically slow preadipocyte replication and differentiation in corpulent rats

1990 ◽  
Vol 258 (2) ◽  
pp. E368-E376 ◽  
Author(s):  
G. Shillabeer ◽  
J. M. Forden ◽  
J. C. Russell ◽  
D. C. Lau
Keyword(s):  
Adipose Tissue ◽  
Doubling Time ◽  
Tissue Growth ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Sprague Dawley ◽  
Induced Differentiation ◽  
Sd Rats

We have investigated the in vitro rate of replication and differentiation of preadipocytes derived from lean (+/+) and obese (cp/cp) male JCR:LA-corpulent (cp) rats in an attempt to identify mechanisms that regulate adipose tissue growth. Cp/cp rats were twofold heavier than age-matched lean rats by 9-10 mo. Cp/cp-derived preadipocytes demonstrated an inherently slower rate of replication than +/+ preadipocytes (population doubling time: cp/cp 52.3 +/- 9.6 h vs. +/+ 19.7 +/- 1.6 h), although the preadipocyte pool in the cp/cp was significantly greater. Cp/cp preadipocytes were resistant to hormonally induced differentiation (19.9 +/- 9.4% of cells accumulated lipid) but differentiated when cocultured with mature adipocytes to the same extent as preadipocytes derived from Sprague-Dawley (SD) rats (cp/cp 48.4 +/- 15.2% vs. SD 52.2 +/- 11.9%). In contrast, SD preadipocytes did not differentiate in response to mature adipocytes from +/+ rats (13.8 +/- 5.2%). Our observations suggest that preadipocyte replication and maturation may not be controlled in a coordinated manner.

scholarly journals The Influence of Aging on the Regenerative Potential of Human Adipose Derived Mesenchymal Stem Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Monika Marędziak ◽  
Krzysztof Marycz ◽  
Krzysztof A. Tomaszewski ◽  
Katarzyna Kornicka ◽  
Brandon Michael Henry
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipogenic Differentiation ◽  
Population Doubling ◽  
Osteogenic Potential ◽  
Population Doubling Time ◽  
Joint Diseases ◽  
Differentiation Potential ◽  
Age Related

Tissue regeneration using human adipose derived mesenchymal stem cells (hASCs) has significant potential as a novel treatment for many degenerative bone and joint diseases. Previous studies have established that age negatively affects the proliferation status and the osteogenic and chondrogenic differentiation potential of mesenchymal stem cells. The aim of this study was to assess the age-related maintenance of physiological function and differentiation potential of hASCs in vitro. hASCs were isolated from patients of four different age groups: (1) >20 years (n=7), (2) >50 years (n=7), (3) >60 years (n=7), and (4) >70 years (n=7). The hASCs were characterized according to the number of fibroblasts colony forming unit (CFU-F), proliferation rate, population doubling time (PDT), and quantified parameters of adipogenic, chondrogenic, and osteogenic differentiation. Compared to younger cells, aged hASCs had decreased proliferation rates, decreased chondrogenic and osteogenic potential, and increased senescent features. A shift in favor of adipogenic differentiation with increased age was also observed. As many bone and joint diseases increase in prevalence with age, it is important to consider the negative influence of age on hASCs viability, proliferation status, and multilineage differentiation potential when considering the potential therapeutic applications of hASCs.

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