scholarly journals Prospective Isolation and Characterization of Chondroprogenitors from Human Chondrocytes Based on CD166/CD34/CD146 Surface Markers

Cartilage ◽  
2021 ◽  
pp. 194760352110424
Author(s):  
Elizabeth Vinod ◽  
Kawin Padmaja ◽  
Abel Livingston ◽  
Jithu Varghese James ◽  
Soosai Manickam Amirtham ◽  
...  

Purpose Chondrocytes, isolated from articular cartilage, are routinely utilized in cell-based therapeutics for the treatment of cartilage pathologies. However, restoration of the biological tissue faces hindrance due to the formation of primarily fibrocartilaginous repair tissue. Chondroprogenitors have been reported to display superiority in terms of their chondrogenic potential and lesser proclivity for hypertrophy. In line with our recent results, comparing chondroprogenitors and chondrocytes, we undertook isolation of progenitors from the general pool of chondrocytes, based on surface marker expression, namely, CD166, CD34, and CD146, to eliminate off-target differentiation and generate cells of stronger chondrogenic potential. This study aimed to compare chondrocytes, chondroprogenitors, CD34−CD166+CD146+ sorted chondrocytes, and CD34−CD166+CD146− sorted chondrocytes. Methods Chondrocytes obtained from 3 human osteoarthritic knee joints were subjected to sorting, to isolate CD166+ and CD34− subsets, and then were further sorted to obtain CD146+ and CD146− cells. Chondrocytes and fibronectin adhesion-derived chondroprogenitors served as controls. Assessment parameters included reverse transcriptase polymerase chain reaction for markers of chondrogenesis and hypertrophy, trilineage differentiation, and total GAG/DNA content. Results Based on gene expression analysis, CD34−CD166+CD146+ sorted chondrocytes and chondroprogenitors displayed comparability and significantly higher chondrogenesis with a lower tendency for hypertrophy when compared to chondrocytes and CD34−CD166+CD146− sorted chondrocytes. The findings were also reiterated in multilineage potential differentiation with the 146+ subset and chondroprogenitors displaying lower calcification and chondroprogenitors displaying higher total GAG/DNA content compared to chondrocytes and 146− cells. Conclusion This unique progenitor-like population based on CD34−CD166+CD146+ sorting from chondrocytes exhibits efficient potential for cartilage repair and merits further evaluation for its therapeutic application.

2020 ◽  
Vol 10 (1) ◽  
pp. 8-16
Author(s):  
Xibin Liu ◽  
Shuang Zhang ◽  
Weijun Guan ◽  
Dong Zheng

Hepatic mesenchymal stem cells (HMSCs) are multipotent stem cells that is a vital part of the regeneration of hepatocytes after injury. In this study, HMSCs were isolated in embryonic livers from of 12-day-old chick embryo using collagenase, and the primary HMSCs were sub-cultured to passage. The protein markers of HMSCs, namely CD71, CD29 and CD44, were tested with immunofluorescence and Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The proliferation of HMSCs in different passages was detected using growth curve, which shown a typically sigmoidal. And then, the pluripotent of HMSCs was analyzed, the results showed that HMSCs could directly induce to differentiate into neural-like cells, adipocytes, and osteoblasts. Our data illustrated that the chick HMSCs have same characteristics to those obtained from other species. The capacity of these cells for multilineage differentiation shows promise for many potential applications.


2013 ◽  
Vol 34 (1) ◽  
pp. 26-35
Author(s):  
Consolatha J.N. Mhaiki ◽  
Enock Masanja ◽  
Jamidu H.Y. Katima ◽  
Gunaraths Rajarao ◽  
Gunnel Dalhammar

Investigation of microorganisms naturally acclimatized to Agave hybrid H 11648 (sisal bole rot) was conducted, with the aim of isolating and characterizing Aspergillus niger strains for industrial use. Microorganism were identified morphologically and then confirmation made by polymerase chain reaction (PCR). Results showed the existence of four major groups, listed in order of abundances as follows; Aspergilli (36.0±0.8) %, Penicillin (28.0±0.1) %, Yeast (15.0±1.6) %and Fusarium (10.0±0.12) %. The main groups of Aspergilli strains were A. nidulans, A. tamari and A. niger in ratios (3:2:2), respectively. Several endo-spore forming non-enteric gram (-) rods and coccid bacteria identified by API20 NE identification systemincluded,Brevundimonas diminuta sp, Shewanella putrefaciens sp, Brevundimonas vesicularis sp and Pasteurella sp. Results showed that sisal bole rot stems hosts a high bio-diversity of microorganism species other than A. niger. Exploitation of the individual strains is recommended. This could eventually produce strains forprecursors of industrially and therapeutically metabolites.


Genome ◽  
2009 ◽  
Vol 52 (12) ◽  
pp. 1037-1039 ◽  
Author(s):  
Walter Durka

Swamp spurge ( Euphorbia palustris , Euphorbiaceae) is a large perennial species of wet grassland and swamps. Its natural habitats are fragmented and isolated both naturally and owing to habitat destruction by human activity. Thus the species is endangered and legally protected in Germany. This report describes seven novel polymorphic microsatellite loci that will be helpful to characterize genetic variation and to analyze the population genetic structure and levels of gene flow within and among populations. All loci were amplified within one multiplex polymerase chain reaction for two populations, yielding between 3 and 13 alleles per locus and high levels of heterozygosity. Trans-species amplification is reported for four Euphorbia species.


1998 ◽  
Vol 330 (2) ◽  
pp. 623-626 ◽  
Author(s):  
Wieke C. C. de BRUIN ◽  
René H. M. te MORSCHE ◽  
J. M. Muriel WAGENMANS ◽  
C. Jeroen ALFERINK ◽  
J. Alan TOWNSEND ◽  
...  

Screening of a genomic mouse DNA library with a glutathione S-transferase class mu cDNA probe resulted in the identification of mGSTM5, a novel member of the murine glutathione S-transferase class mu gene family. Here we present the sequence of the promoter region, the exon-intron organization of the gene and the isolation and characterization of its complete cDNA. Conceptual translation of the cDNA sequence revealed that several amino acid positions have been changed in ‘invariant’ mu class signature sequences in mGSTM5. Reverse transcriptase polymerase chain reaction using gene specific primers revealed that mGSTM5 is uniquely expressed in mouse liver, stomach and small intestine.


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