Limits...
Gene expression profile of epithelial cells and mesenchymal cells derived from limbal explant culture.

Polisetti N, Agarwal P, Khan I, Kondaiah P, Sangwan VS, Vemuganti GK - Mol. Vis. (2010)

Bottom Line: The microarray data was validated by using Realtime and semiquntitative reverse transcription polymerase chain reaction.We have also observed similar and differential gene expression between MC-L and MSC-BM.This study represents the first extensive gene expression analysis of limbal explant culture derived epithelial and mesenchymal cells and as such reveals new insight into the biology, ontogeny, and in vivo function of these cells.

View Article: PubMed Central - PubMed

Affiliation: C-TRACER, Hyderabad Eye Research Foundation, LV Prasad Eye Institute, Hyderabad, India.

ABSTRACT

Purpose: Limbal stem cell deficiency is a challenging clinical problem and the current treatment involves replenishing the depleted limbal stem cell (LSC) pool by either limbal tissue transplantation or use of cultivated limbal epithelial cells (LEC). Our experience of cultivating the LEC on denuded human amniotic membrane using a feeder cell free method, led to identification of mesenchymal cells of limbus (MC-L), which showed phenotypic resemblance to bone marrow derived mesenchymal stem cells (MSC-BM). To understand the transcriptional profile of these cells, microarray experiments were carried out.

Methods: RNA was isolated from cultured LEC, MC-L and MSC-BM and microarray experiments were carried out by using Agilent chip (4x44 k). The microarray data was validated by using Realtime and semiquntitative reverse transcription polymerase chain reaction.

Results: The microarray analysis revealed specific gene signature of LEC and MC-L, and also their complementary role related to cytokine and growth factor profile, thus supporting the nurturing roles of the MC-L. We have also observed similar and differential gene expression between MC-L and MSC-BM.

Conclusions: This study represents the first extensive gene expression analysis of limbal explant culture derived epithelial and mesenchymal cells and as such reveals new insight into the biology, ontogeny, and in vivo function of these cells.

Show MeSH

Related in: MedlinePlus

Morphological features of mesenchymal cells of limbus. Limbal explant cultures having epithelial (E) and mesenchymal cells (S; 200×; A). Cell sphere formation in the MC-L cultures giving impression of embryoid body formation (200×; B). Spindle shaped morphology of MC-L forming colonies (200×; C). Culture of MC-L showing both spindle shaped and broad flattened cells (arrows; 200×; D)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2903463&req=5

f1: Morphological features of mesenchymal cells of limbus. Limbal explant cultures having epithelial (E) and mesenchymal cells (S; 200×; A). Cell sphere formation in the MC-L cultures giving impression of embryoid body formation (200×; B). Spindle shaped morphology of MC-L forming colonies (200×; C). Culture of MC-L showing both spindle shaped and broad flattened cells (arrows; 200×; D)

Mentions: Spindle cell cultures were established from extended limbal explant cultures after 2 to 3 weeks of culture. Under a phase contrast microscope the cells appeared fibroblastic, elongated, and spindle shaped and few cells were large and flat with a single nucleus. These cells demonstrated the ability to form colonies with the occasional cell sphere formation giving the impression of embryoid bodies (Figure 1).


Gene expression profile of epithelial cells and mesenchymal cells derived from limbal explant culture.

Polisetti N, Agarwal P, Khan I, Kondaiah P, Sangwan VS, Vemuganti GK - Mol. Vis. (2010)

Morphological features of mesenchymal cells of limbus. Limbal explant cultures having epithelial (E) and mesenchymal cells (S; 200×; A). Cell sphere formation in the MC-L cultures giving impression of embryoid body formation (200×; B). Spindle shaped morphology of MC-L forming colonies (200×; C). Culture of MC-L showing both spindle shaped and broad flattened cells (arrows; 200×; D)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2903463&req=5

f1: Morphological features of mesenchymal cells of limbus. Limbal explant cultures having epithelial (E) and mesenchymal cells (S; 200×; A). Cell sphere formation in the MC-L cultures giving impression of embryoid body formation (200×; B). Spindle shaped morphology of MC-L forming colonies (200×; C). Culture of MC-L showing both spindle shaped and broad flattened cells (arrows; 200×; D)
Mentions: Spindle cell cultures were established from extended limbal explant cultures after 2 to 3 weeks of culture. Under a phase contrast microscope the cells appeared fibroblastic, elongated, and spindle shaped and few cells were large and flat with a single nucleus. These cells demonstrated the ability to form colonies with the occasional cell sphere formation giving the impression of embryoid bodies (Figure 1).

Bottom Line: The microarray data was validated by using Realtime and semiquntitative reverse transcription polymerase chain reaction.We have also observed similar and differential gene expression between MC-L and MSC-BM.This study represents the first extensive gene expression analysis of limbal explant culture derived epithelial and mesenchymal cells and as such reveals new insight into the biology, ontogeny, and in vivo function of these cells.

View Article: PubMed Central - PubMed

Affiliation: C-TRACER, Hyderabad Eye Research Foundation, LV Prasad Eye Institute, Hyderabad, India.

ABSTRACT

Purpose: Limbal stem cell deficiency is a challenging clinical problem and the current treatment involves replenishing the depleted limbal stem cell (LSC) pool by either limbal tissue transplantation or use of cultivated limbal epithelial cells (LEC). Our experience of cultivating the LEC on denuded human amniotic membrane using a feeder cell free method, led to identification of mesenchymal cells of limbus (MC-L), which showed phenotypic resemblance to bone marrow derived mesenchymal stem cells (MSC-BM). To understand the transcriptional profile of these cells, microarray experiments were carried out.

Methods: RNA was isolated from cultured LEC, MC-L and MSC-BM and microarray experiments were carried out by using Agilent chip (4x44 k). The microarray data was validated by using Realtime and semiquntitative reverse transcription polymerase chain reaction.

Results: The microarray analysis revealed specific gene signature of LEC and MC-L, and also their complementary role related to cytokine and growth factor profile, thus supporting the nurturing roles of the MC-L. We have also observed similar and differential gene expression between MC-L and MSC-BM.

Conclusions: This study represents the first extensive gene expression analysis of limbal explant culture derived epithelial and mesenchymal cells and as such reveals new insight into the biology, ontogeny, and in vivo function of these cells.

Show MeSH
Related in: MedlinePlus