June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Mesenchymal stem cells for bilateral limbal stem cell deficiency: Expression profile of different markers and status of transdifferentiation into corneal epithelial cells
Author Affiliations & Notes
  • Sachin Shukla
    Centre for Ocular Regeneration, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
  • D Ram Reddy
    Centre for Ocular Regeneration, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
  • Madhuri Batta
    Centre for Ocular Regeneration, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
  • Vivek Singh
    Centre for Ocular Regeneration, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
  • Virender Singh Sangwan
    Centre for Ocular Regeneration, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
  • Footnotes
    Commercial Relationships   Sachin Shukla, None; D Reddy, None; Madhuri Batta, None; Vivek Singh, None; Virender Sangwan, None
  • Footnotes
    Support  DST-INSPIRE Faculty Grant (IFA14-LSBM-104), Department of Science and Technology, Govt. of India
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1379. doi:
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      Sachin Shukla, D Ram Reddy, Madhuri Batta, Vivek Singh, Virender Singh Sangwan; Mesenchymal stem cells for bilateral limbal stem cell deficiency: Expression profile of different markers and status of transdifferentiation into corneal epithelial cells. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1379.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Patients with bilateral deficiency of limbal stem cells present a unique set of challenges to the clinician due to the clinical presentation, underlying causes, adnexal status, and lack of an autologous source of limbal stem cells; altogether making it difficult to devise a successful treatment. We are exploring mesenchymal stem cells derived from the dental pulp and adipose tissue to test the hypothesis that they can be used as better alternatives for treating the bilateral limbal stem cell deficiency due to their properties of immunomodulation and multipotent differentiation.

Methods : The human MSCs derived from the dental pulp (DPSCs) and adipose tissues (ADSCs) were procured from the Stemade Biotech Pvt. Ltd. (Mumbai, India) and Thermo Fisher Scientific Inc. (USA), respectively. Rabbit corneal epithelial cell line (SIRC) was procured from the National Centre for Cell Science, Pune, India and was used as a control. The cells were cultured under standard culture conditions (37 degree Celsius and 5% CO2). The DPSCs and SIRC were cultured in the DMEM supplemented with 10% fetal bovine serum, whereas ADSCs were cultured in MesenPRO RS™ media (Gibco™, Thermo Fisher Scientific Inc., USA), respectively. Expression of mesenchymal and corneal epithelial markers was studied through semiquantitative RT-PCR and immunofluorescence. Induction medium or co-culture methods were used for studying the transdifferentiation.

Results : Immunofluorescence analysis revealed the expression of CD90, CD105, and Vimentin in both the DPSCs and ADSCs; CD14 could not be detected in both of them whereas CD34 was detected only in ADSCs. The DPSCs also showed the expression of stem cell markers ABCG2 and p63α, whereas SIRC showed the expression of corneal epithelial markers PAX6 and CK3/12.

Conclusions : The results support the mesenchymal origin of both the dental pulp- and adipose-derived MSCs and present a comparative analysis of expression of mesenchymal, epithelial, and stem cell markers in these cells.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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