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Isolation, culture, purification and ultrastructural investigation of cardiac telocytes.

Li YY, Zhang S, Li YG, Wang Y - Mol Med Rep (2016)

Bottom Line: Furthermore, immunofluorescence demonstrated that almost all the sorted TCs expressed vimentin, a marker of TCs.Moreover, electron micrographs showed typical TCs based on their ultrastructural features.Using this method, we developed a reproducible protocol for the isolation and purification of cardiac TCs from rat hearts, which yielded TCs with typical characteristics.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Diseases, Xinhua Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai 200092, P.R. China.

ABSTRACT
Telocytes (TCs), a novel type of stromal cells, are crucial to cardiac renovation and regeneration. To dissect the pathophysiological effects of cardiac TCs in heart disease, it is essential to develop an effective method to isolate, culture, purify and characterize these cells. In the present study, cardiac TCs were isolated from the hearts of rats by enzymatic digestion. Histology and CD34/PDGFRα expression by flow cytometric assay were used to characterize the cultured cardiac TCs, which were purified by flow cytometric sorting and confirmed by immunofluorescence and electron microscopy. Typical TCs were observed in primary culture, with these exhibiting typical fusiform cell bodies with long moniliform telopodes. Based on flow cytometric sorting with antibodies to CD34 and PDGFRα, there was a substantial increase in the purity of cardiac TCs. Furthermore, immunofluorescence demonstrated that almost all the sorted TCs expressed vimentin, a marker of TCs. Moreover, electron micrographs showed typical TCs based on their ultrastructural features. Using this method, we developed a reproducible protocol for the isolation and purification of cardiac TCs from rat hearts, which yielded TCs with typical characteristics.

No MeSH data available.


Related in: MedlinePlus

Transmission electron micrograph of a TC. (A) TCs had a small oval body, mostly occupied by the nucleus, encircled by a small amount of cytoplasm. (B) The structures in the cytoplasm were: m, mitochondria; li, lipid droplets; g, Golgi apparatus; er, endoplasmic reticulum; and NC, nucleus. (C) Higher magnification of a telopode. TC, telocyte.
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f4-mmr-14-02-1194: Transmission electron micrograph of a TC. (A) TCs had a small oval body, mostly occupied by the nucleus, encircled by a small amount of cytoplasm. (B) The structures in the cytoplasm were: m, mitochondria; li, lipid droplets; g, Golgi apparatus; er, endoplasmic reticulum; and NC, nucleus. (C) Higher magnification of a telopode. TC, telocyte.

Mentions: Using electron microscopy, we noted that TCs had a small oval body, mostly occupied by the nucleus and encircled by a small amount of cytoplasm (Fig. 4A). Additionally, we observed that the cytoplasm was filled with mitochondria, lipid droplets, a small Golgi apparatus, in addition to elements of smooth and rough endoplasmic reticulum (Fig. 4B). Furthermore, higher magnification images of Tps were obtained (Fig. 4C). As one of the most striking features of TCs, the connections were organized through homocellular junctions. Figure 5A shows that tight contacts (atypical junctions) occurred between Tps and the TC cell body. Apart from the homocellular junctions, TCs have been demonstrated to serve an important role in cellular communication in the heart by releasing extracellular vesicles (EVs) (Fig. 5B–D). Different types of EVs were observed: Exosomes, mainly as intraluminal vesicles filled with multivesicular bodies (Fig. 5B); ectosomes, released from the plasma membrane of the cell body and Tps (Fig. 5C); and clusters of endomembrane vesicles encircled by the plasma membrane, termed multivesicular cargo (Fig. 5D).


Isolation, culture, purification and ultrastructural investigation of cardiac telocytes.

Li YY, Zhang S, Li YG, Wang Y - Mol Med Rep (2016)

Transmission electron micrograph of a TC. (A) TCs had a small oval body, mostly occupied by the nucleus, encircled by a small amount of cytoplasm. (B) The structures in the cytoplasm were: m, mitochondria; li, lipid droplets; g, Golgi apparatus; er, endoplasmic reticulum; and NC, nucleus. (C) Higher magnification of a telopode. TC, telocyte.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940097&req=5

f4-mmr-14-02-1194: Transmission electron micrograph of a TC. (A) TCs had a small oval body, mostly occupied by the nucleus, encircled by a small amount of cytoplasm. (B) The structures in the cytoplasm were: m, mitochondria; li, lipid droplets; g, Golgi apparatus; er, endoplasmic reticulum; and NC, nucleus. (C) Higher magnification of a telopode. TC, telocyte.
Mentions: Using electron microscopy, we noted that TCs had a small oval body, mostly occupied by the nucleus and encircled by a small amount of cytoplasm (Fig. 4A). Additionally, we observed that the cytoplasm was filled with mitochondria, lipid droplets, a small Golgi apparatus, in addition to elements of smooth and rough endoplasmic reticulum (Fig. 4B). Furthermore, higher magnification images of Tps were obtained (Fig. 4C). As one of the most striking features of TCs, the connections were organized through homocellular junctions. Figure 5A shows that tight contacts (atypical junctions) occurred between Tps and the TC cell body. Apart from the homocellular junctions, TCs have been demonstrated to serve an important role in cellular communication in the heart by releasing extracellular vesicles (EVs) (Fig. 5B–D). Different types of EVs were observed: Exosomes, mainly as intraluminal vesicles filled with multivesicular bodies (Fig. 5B); ectosomes, released from the plasma membrane of the cell body and Tps (Fig. 5C); and clusters of endomembrane vesicles encircled by the plasma membrane, termed multivesicular cargo (Fig. 5D).

Bottom Line: Furthermore, immunofluorescence demonstrated that almost all the sorted TCs expressed vimentin, a marker of TCs.Moreover, electron micrographs showed typical TCs based on their ultrastructural features.Using this method, we developed a reproducible protocol for the isolation and purification of cardiac TCs from rat hearts, which yielded TCs with typical characteristics.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Diseases, Xinhua Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai 200092, P.R. China.

ABSTRACT
Telocytes (TCs), a novel type of stromal cells, are crucial to cardiac renovation and regeneration. To dissect the pathophysiological effects of cardiac TCs in heart disease, it is essential to develop an effective method to isolate, culture, purify and characterize these cells. In the present study, cardiac TCs were isolated from the hearts of rats by enzymatic digestion. Histology and CD34/PDGFRα expression by flow cytometric assay were used to characterize the cultured cardiac TCs, which were purified by flow cytometric sorting and confirmed by immunofluorescence and electron microscopy. Typical TCs were observed in primary culture, with these exhibiting typical fusiform cell bodies with long moniliform telopodes. Based on flow cytometric sorting with antibodies to CD34 and PDGFRα, there was a substantial increase in the purity of cardiac TCs. Furthermore, immunofluorescence demonstrated that almost all the sorted TCs expressed vimentin, a marker of TCs. Moreover, electron micrographs showed typical TCs based on their ultrastructural features. Using this method, we developed a reproducible protocol for the isolation and purification of cardiac TCs from rat hearts, which yielded TCs with typical characteristics.

No MeSH data available.


Related in: MedlinePlus