End-Stage liver Disease (ESLD) is a major healthcare-related problem growing every year in India and worldwide. There are only two clinical options available as treatment modality, which include liver transplant and primary hepatocytes transplant. Both options have limitations due to inadequate availability of donors. The development of a transplantable artificial liver organ can offer a solution to this problem. In this regard, decellularized liver organ biomatrix (DLEM) offers a relevant biological and clinical option. Repopulation of DLEM with hepatocytes can provide a functional tissue with transplantation potential. However, the limitation in availability of primary hepatocytes and derivation of hepatocytes like cells (HLC) from stem cells is a cost-intensive, long-duration protocol (between 21 to 28 days). This limits its large-scale production and use for whole organ development. To address these problems, we have developed differentiation strategies to derive functional hepatocytes and hepatic tissue using a direct differentiation [1] and stage-specific differentiation approach [2].
In our first approach, we tested the hepatogenic capability of hyperbilirubinemia patient derived sera. This process resulted in the development of functional HLC in 7 days [1]. In our second approach, we used a combination of small molecule activators and inhibitors in a sequential manner to derived HLC in four stages mimicking liver development in utero. This process resulted in the development of functional HLC in 14 days and exhibited molecular and functional characteristics similar to growth factor-based differentiation protocol which takes 23-28 days for hepatocytes like cell derivation [2]. For hepatic tissue development, we used the optimized differentiation strategy to develop functional hepatic tissue using mesenchymal stem cell (MSC) repopulated human decellularized liver biomatrix (DLEM) [2]. The derived HLC was transplanted in an acute liver injury model of rat to study the improvement in liver function of the model and their survival. These strategies can be useful for both cell-based therapy and transplantable hepatic tissue-based liver tissue engineering and regeneration for potential in vitro hepatic model development and clinical application.

Publications:
[1] Santosh Gupta, Akriti Sharma, Sugan Paneerselvan, Sangeeta Kandhoi, Bamadeb Patra, Dillip Kumar Bishi, Kumar Narayanaswamy, Rama Shanker Verma. Human Derived Hepatogenic Serum Induced Hepatic Specified Cells for Liver Injury Treatment in rats. Biomaterials. (Under review, Xenotransplantation)
[2] Santosh Gupta, Akriti Sharma, Mohamed Rela, Rama Shanker Verma. Chemically Induced Hepatocytes like Cells and Hepatic tissue for Acute Liver Injury Treatment. Journal of Tissue Engineering and Regenerative Medicine. (Under review, Biomaterials)
[3].Santosh Gupta, Akriti Sharma, Archana S, Rama Shanker Verma, Mesenchymal Stem Cells for Cardiac Regeneration: From Differentiation to Cell Delivery, Stem Cell Review Reports, 2021, https://doi.org/10.1007/s12015-021-10168-0.
[4] Santosh Gupta, Akriti Sharma, Rama Shanker Verma. Polymers in biosensor devices for cardiovascular applications. Current Opinion in Biomedical Engineering 2020 Mar 13;69-75. https://doi.org/10.1016/j.cobme.2019.10.002
[5] Santosh Gupta, Akriti Sharma, Vasantha Kumar J, Vineeta Sharma, Piyush Kumar Gupta, Rama Shanker Verma. Meniscal Tissue Engineering via 3D Printed PLA Monolith with Carbohydrate based Self- Healing Interpenetrating Network Hydrogel. International Journal of Biological Macromolecules. 2020, 162,1358-1371. https://doi.org/10.1016/j.ijbiomac.2020.07.238.
[6] Piyush K Gupta, Surya Kant Tripathi, SreenathPappuru, Shiva ChanderChabattula, Kavitha Govarthanan, Santosh Gupta, Bijesh K Biswal, Debashis Chakraborty, Rama S Verma. (2019). Metal-free semi-aromatic polyester as nanodrug carrier: A novel tumor-targeting drug delivery vehicle for potential clinical application. Material Science and Engineering C: Materials for Biological Application. https://doi.org/10.1016/j.msec.2019.110285
[7] Piyush Kumar Gupta, Santosh Gupta, SreenathPappuru, Siva ChanderChabattula, Debashis Chakraborty, Rama Shanker Verma. Enhancing the anti-cancer therapeutic efficacy by optimizing molecular weight of metal-free fully alternating semi-aromatic polyester as nano-drug carriers. Journal of Drug Delivery Science and Technology, Volume 51, June 2019, Pages 101-114
[8] Gupta PK, Pappuru S, Gupta S, Patra B, Chakraborty D, Verma RS. Self-assembled dual-drug loaded core-shell nanoparticles based on metal-free fully alternating polyester for cancer theranostics. Mater Sci Eng C Mater Biol Appl. 2019 Aug; 101:448-463.
[9] Diptendu Chatterjee, Piyush Kumar Gupta, Santosh Gupta, and Rama Shanker Verma. (2018) Fanconi Anemia: A Rare Genetic Disorder in Man JIAS (Journal of Indian Anthropological Society)53:265-266(2018).
[10] Gupta S, Sharma V, Verma RS. Recent Advances in Induced Pluripotent Stem Cell (iPSC) based Therapeutics. Journal of Stem Cell Research & Therapeutics - 2017;3(3):263-270. DOI: 10.15406/jsrt.2017.03.00100
[11] Gupta S, Gupta PK, Dharanivasan G, Verma RS. Current prospects and challenges of nanomedicine delivery in prostate cancer therapy. Nanomedicine (Lond). 2017 Dec;12(23):2675-2692. doi: 10.2217/nnm-2017-0236.