Lalita Limaye

Area of Research

Cryopreservation, ex vivo expansion and differentiation of Hematopoietic , Mesenchymal and Induced pluripotent stem cells.

Research Projects

  1. Ex vivo expansion and cryopreservation of human umbilical cord blood stem cells.

  2. In vitro generation, characterization and cryopreservation of Megakaryocytes and platelets from Hematopoietic stem cells.

  3. In vitro generation, characterization and cryopreservation of dendritic cells from hematopoietic stem cells.

  4. Development of MSCs from Umbilical cord blood and their differentiation to neural cells.

  5. Studying the Effect of oral feeding of nutraceuticals belonging to the class of polyunsaturated fatty acids on hematopoiesis and Thrombopoiesis of mice?

  6. Studies on generation of induced pluripotent stem cells from umbilical cord tissue derived adult stem cells.

  7. In vitro generation of RBCs/Nutrophils from HSCs

  8. Research Details

    Ex vivo expansion and cryopreservation of human umbilical cord blood stem cells:

    In-vitro expansion of CD34+ cells is being extensively attempted to overcome the limitations of the small sample size especially of cord blood and pediatric marrow harvests. Optimal storage of these expanded cells is also crucial. There is a possibility that during such in-vitro manipulations the cells undergo apoptosis. To date a body of evidence has been compiled which attests to the hypothesis that cold shock can induce apoptosis. Thus detection of the level of apoptosis during preservation and expansion and to develop ways and means to prevent it is an important aspect of hematopoietic cell research. We have used anti-apoptotic agents as supplements in media for in vitro expansion and cryopreservation of expanded CD34+ cells and see that they reduce apoptosis and enhance expansion. We are now focusing on use of anti-apoptotic agents for expansion of HSCs in a co-culture system.

    In vitro generation, characterization and cryopreservation of Megakaryocytes and platelets from Hematopoietic stem cells:

    Umbilical cord blood provides an alternative source of hematopoietic progenitor cells for transplantation. However prolonged thrombocytopenia remains a major obstacle due to the lower numbers of megakaryocyte progenitor cells and their subsequent delayed engraftment. Efforts currently are underway to expand the number of CB cells and MK progenitor cells ex-vivo, which may facilitate platelet production during post-transplant nadir and decrease the time of thrombocytopenia. We found that polyunsaturated fatty acids (PUFAs) have a beneficial effect on megakaryocyte generation. We are now trying to understand the molecular and cellular events occurring during platelet biogenesis with special reference to polyunsaturated fatty acids.

    In vitro generation ,characterization and cryopreservation of Dendritic cells from hematopoietic stem cells:

    Dendritic cells form a heterogeneous population of cells capable of stimulating naive T cells and initiating primary immune response. This well-known function of DCs has offered the possibility of developing clinical protocol for their use in immunotherapy to tumors. The total numbers of DCs available for immunotherapy remains limited .DCs have been shown to be generated from CD34+ cells from BM, UCB and G CSF mobilized PBSC. Ex-vivo generation of this cell type and its efficient cryopreservation will have direct application in the clinics. We have optimized protocols for generation and cryopreservation of DCs from Cord blood derived CD34+ cells. We are now carrying out in depth studies on functional aspects of these UCB derived DCs.


    Development of MSCs from Umbilical cord blood and their differentiation to neural cells:

    Due to their self-renewal and differentiation ability stem cells are being largely used in cellular replacement therapies particularly in neurodegenerative diseases. Most clinical applications of stem cells rely on development of in vitro protocols for their efficient expansion and differentiation. Although a number of investigators all over the world are involved in this type of research, the knowledge and methods of differentiating stem cells into specialized cell types is still limited. MSCs (Mesenchymal stem cells) have been derived from different sources like Bone marrow, peripheral blood, adipose tissue and umbilical cord blood. It has been described that multipotent MSCs are capable of differentiating into a variety of cell types from different germ layers.

    In the present proposal we are attempting to standardize culture conditions for generation of MSCs from umbilical cord blood. Once these methods are standardized then we will focus on their differentiation to neural cells. Thus this project will help us in gaining knowledge of the molecular and cellular mechanisms involved in differentiation of cord blood/cord tissue derived MSCs to neural cells. This type of study will find applications in future, in cell replacement therapies in neuronal diseases like Parkinson's disease, Alzheimer disease, spinal cord injuries etc.

    Studying the Effect of oral feeding of nutraceuticals belonging to the class of polyunsaturated fatty acids on hematopoiesis and Thrombopoiesis of mice:

    Diet plays a key role in human health and it has a significant effect on homeostasis. Diet rich in fruits and vegetables lowers the rate of chronic diseases like, cardiovascular disease as well as cancer. Omega-3 (n-3) and omega-6 (n-6) polyunsaturated fatty acids (PUFAs) are important structural and functional components of cell membrane phospholipids. In the present proposal our main goal will be to investigate, whether n-3 and n-6 PUFAs affect hematopoietic stem/progenitor cells (hematopoiesis) and Thrombopoiesis in vivo. Since both n-3 and n-6 PUFAs are commonly classified as functional foods, it is imperative to study the effect of an oral administration of these PUFAs on the Hematopoiesis and Thrombopoiesis of mice.

    Studies on generation of induced pluripotent stem cells from umbilical cord tissue derived adult stem cells:

    Induced pluripotent stem cells (iPSCs) are cells that resemble ESCs that have the property of self renewal and pluripotency. We hypothesize that the adult stem cell source is a much more primitive cell type as compared to somatic cells that could be employed for reprogramming. The advantage of using this cell type is that it might maintain a greater genomic stability and use of lesser number of factors as these cells indigenously express some of the widely known reprogramming genes.Once iPSCs have been generated we would like to further differentiate them into cells of haematopoietic lineage. If the protocols of differentiation of iPSCs to cells of various haematopoietic lineages specifically to HSCs would be standardized it can prove to be a better source as compared to the conventional methods.

    In vitro generation of RBCs and Neutrophils from HSCs:

    In vivo production of red blood cells occurs in the bone marrow niche. HSCs residing in bone marrow respond to cell based as well as soluble signals to commit to the erythroid progenitors. These committed progenitors interact with the cells present in stromal environment that support their differentiation to form mature RBCs .Various reports have been published that mention protocols for ex-vivo generation of RBCs from HSCs by using co-culture system with murine stromal cells and human derived bone marrow mesenchymal stem cells (MSCs). Since murine stromal cells have xenogenic origin they cannot be used for human application. Further, bone marrow harvesting is a painful process. So we planned to use two human derived MSCs isolated from cord and placental tissues which are considered as clinical waste. These MSCs will be used in RBC generation.

    In a similar way, neutrophil generation is gaining attention recently and very few reports regarding their in vitro generation are published . So, there is a need to further optimize the protocol and study the mechanism involved in its generation.Thus in the present project we will focus on in vitro generation of these two cell types from HSCs.

    Technology Developed

    In 1998 the technology of bone marrow cryopreservation was developed and transferred to Armed Forces Medical College (AFMC), Pune. In 1998 the technology of bone marrow cryopreservation was developed and transferred to Armed Forces Medical College (AFMC), Pune.


    Overseas Fellowship

    Recipient of DBT short term overseas fellowship in 2008

    Worked in Dr. Meri Firpo's Lab.,Stem Cell Institute, McGuire Translational research facility, 2001,6th Street S.E. University of Minnesota, Minneapolis, MN 55455 on the project: "To examine the effect of anti-oxidants and membrane stabilizers on efficacy of cryopreservation of human ES cells."


    Intramural Funding : By NCCS

    Extramural funding :

    3 projects (completed) DRDO (Life Science Research Board)
    4 projects (completed) DBT
    1 project (completed) DAE BRNS


    Sophia Fernandes a, Manasi Talwadekar a, Raju Agarwal b,1, Velu Nair c,2, Vaijayanti Kale a, Lalita Limaye Generation and characterization of human iPSC line from CD34+ cells isolated from umbilical cord blood belonging to Indian origin.Accepted in Dec. 2016 in Stem Cell Research.

    Kedar Limbkar, Vaijayanti Kale and Lalita Limaye Oral Feeding With Arachidonic Acid (AA) and Docosahexanoic Acid (DHA) Help in Better Recovery of Haematopoiesis in Sub-lethally Irradiated Mice Biomed Res J 2016;3(2):182-194

    Sapana Jalnapurkar, Shweta Singh, Moirangthem Ranjita Devi, Lalita Limaye and Vaijayanti Kale (2016) Nitric oxide has contrasting age-dependent effects on the functionality of murine hematopoietic stem cells. Stem Cell Research and Therapy. In press.

    Manmohan S. Bajaj, Rohan S. Kulkarni, Suprita S. Ghode, Lalita S. Limaye,Vaijayanti P. Kale Irradiation-induced secretion of BMP4 by marrow cells causes adipogenesis post-myelosuppression.Accepted in Stem cell research.Nov.2016

    Darshana Kadekar, Sonal Rangole, Vaijayanti Kale, Lalita Limaye* PLOS ONE Conditioned Medium from Placental Mesenchymal Stem Cells Reduces Oxidative Stress during the Cryopreservation of Ex Vivo Expanded Umbilical Cord Blood Cells | DOI:10.1371/journal.pone.0165466 October 25, 2016

    Suprita Ghode, Rohan kulkarni, MohanBajaj, Yogesh Shouche,Lalita Limaye,Vaijayanti Kale Neuropilin1 is an important niche component and exerts context-dependent effects on hematopoietic stem cells. Accepted in Sept 2016 in Stem cells and development.

    Manasi Talwadekar a, Sophia Fernandes a, Vaijayanti Kale a and Lalita Limaye,* Valproic acid enhances the neural differentiation of human placenta derived-mesenchymal stem cells in vitro Accepted Journal of Tissue Engineering and regenerative Medicine April 2016

    Shweta Singh, Ranjita Devi Moirangthem, Anuradha Vaidya, Sapana Jalnapurkar, Lalita Limaye and Vaijayanti Kale (2016) Stem Cells doi: 10.1002/stem.2409

    Manasi D. Talwadekar, Vaijayanti P. Kale & Lalita S. Limaye*Placenta-derived mesenchymal stem cells possess better immunoregulatory properties compared to their cord-derived counterparts–a paired sample study.Scientific Reports 5:15784 | DOI: 10.1038/srep15784

    Darshana Kadekar ,Vaijayanti Kale and Lalita S. Limaye *Differential ability of MSCs isolated from placenta and cord as feeders for supporting ex vivo expansion of umbilical cord blood derived CD34+ cells.Stem Cell Research & Therapy (2015) 6:201

    Ankita C.Dhenge, Vaijayanti Kale, Lalita S. Limaye*Enhancement of hematopoiesis and thrombopoiesis by pharmacological compounds valproic acid and all- trans retinoic acid.J.PHARM.SCI.TECH.MGMT. Vol.1 Issue 2 2015

    Jeetendra Kumar, Vaijayanti Kale and Lalita S. Limaye *Umbilical cord blood-derived CD11c+dendritic cells could serve as an alternative allogeneic source of dendritic cells for cancer immunotherapy. Stem Cell Research & Therapy(2015) 6:184 DOI 10.1186/s13287-015-0160-8

    Ranjita Devi Moirangthem, Shweta Singh, Ashwini Adsul, Sapana Jalnapurkar, Lalita S. Limaye and Vaijayanti P. Kale. Hypoxic Niche-Mediated Regeneration of Hematopoiesis in the Engraftment Window Is Dominantly Affected by Oxygen Tension in the Milieu.STEM CELLS AND DEVELOPMENTVolume 24, Number 20, 2015 DOI: 10.1089/scd.2015.0112.

    Shweta Singh, Suprita Ghode, Moirangthem Ranjita Devi, Lalita Limaye and Vaijayanti Kale* Phenotypic and functional characterization of a marrowderived stromal cell line, M210B4 and its comparison with primary marrow stromal cells. Biomedical research journal 2015;2(1):120-133.

    Manmohan Bajaj, Suprita Ghode,Rohan Kulkarni, Lalita Limaye and Vaijayanti Kale "Simvastatinimproveshematopoietic stem cell engraftment by preventing irradiation-induced marrow adipogenesis and radio-protecting the niche cells". Hematologica. 2015 April Hematol.2 015.124750 PMID:25887500.

    Jeetendra Kumar, Rupali Gurav, V P Kale and L S Limaye . Exogenous addition of arachidonic Acid to the culture media enhances the functionality of dendritic cells for their possible use in cancer immunotherapy. PLoS One. 2014 Nov 4;9 (11):e111759.

    Manasi Talwadekar, Darshana Kadekar, Sonal Rangole, Nikhat Firdaus Khan, Vaijayanti Kale and Lalita Limaye *Cultivation and Cryopreservation of Cord Tissue MSCs with Cord Blood AB Plasma.Biomed Res J 2014;1(2):126-136.

    Shabrani NC, Khan NF, Kale VP, Limaye LS. Polyunsaturated fatty acids confer cryoresistance on megakaryocytes generated from cord blood and also enhance megakaryocyte production from cryopreserved cord blood cells. Cytotherapy, 2012 Mar;14(3):366-80. Epub 2012 Jan 17. [Link]

    Sangeetha VM, Kadekar D, Kale VP, Limaye LS. Pharmacological inhibition of caspase and calpain proteases: a novel strategy to enhance the homing responses of cord blood HSPCs during expansion. PLoS One 2012; 7(1):e29383. Epub 2012 Jan 3. [Link]

    Monika B. Sharma, Lalita S. Limaye, and Vaijayanti P. Kale.Mimicking the functional hematopoietic stem cell niche in vitro:recapitulation of marrow physiology by hydrogel-based three-dimensional cultures of mesenchymal stromal cells Haematologica 2011 [Epub ahead of print] 96:xxx doi:10.3324/haematol.2011.050500. [Link]

    Bajaj M, Hinge A, Limaye LS, Gupta RK, Surolia A, Kale VP,Mannose-binding dietary lectins induce adipogenic differentiation of the marrow-derived mesenchymal cells via an active insulin-like signaling mechanism. Glycobiology. 2010 Nov 23. [Link]

    Sangeetha V.M, Vaijayanti P Kale, Lalita S Limaye
    Expansion of cord blood CD34+ cells in presence of zVADfmk and zLLYfmk improved their in vitro functionality and in vivo engraftment in NOD/SCID mouse.PLoS ONE Aug 2010, Vol. 5 Issue 8 [Link]

    Balan S, Kale VP, Limaye LS
    A large number of mature and functional Dendritic Cells can be efficiently generated from umbilical cord blood-derived mononuclear cells by a simple two step culture method.Transfusion March 2010 PMID:20497510 [Link]

    Ashwini S. Hinge,Lalita S. Limaye, Avadhesha Surolia, and Vaijayanti P. Kale
    In vitro protection of umbilical cord blood derived primitive hematopoietic stem progenitor cell pool by mannose-specific lectins via antioxidant mechanisms. Transfusion March 2010 PMID 20412533 [Link]

    Nikhat Firdaus A. Siddiqui, Namrata C. Shabrani, Vaijayanti P.Kale andLalita S.Limaye*
    Enhanced generation of megakaryocytes from UCB-derived CD34+ cells expanded in the presence of two nutraceuticals-Docosahexanoic acid and Arachidonic acid as supplements to the cytokine containing medium. Cytotherapy 2010 PMID: 2023022.[Link]

    Hinge A, Bajaj M, Limaye L,Surolia A, Kale VP.
    Oral administration of insulin receptor-interacting lectins leads to an enhancement in the hematopoietic stem and progenitor cell pool of mice. Stem Cells Dev. 2009 Jul 6. PMID: 19580456.[Citation] [Link]

    Balan S, Kale VP, Limaye LS.
    A simple two-step culture system for the large-scale generation of mature and functional dendritic cells from umbilical cord blood CD34+ cells. Transfusion 2009 May 27. PMID: 19497054.[Citation] [Link]

    Invited review on "The in-vitro Expansion Protocol: Current Status" in The Bio Medic, Published by Reliance Life Sciences,2, Sept. 2007, 28-30

    Lalita M. Sasnoor, Vaijayanti P. Kale and Lalita S. Limaye.
    "Cryopreservation of mouse bone marrow: Prevention of apoptosis as a possible mechanism of improved protection of cells by catalase and trehalose as additives in conventional freezing medium",Transplantation Nov 2005 (vol.80,no.9) 1251-1260 [Citation] [Link]

    Lalita M. Sasnoor, Vaijayanti P. Kale and Lalita S. Limaye.
    "A combination of catalase and trehalose as additives in the conventional freezing medium results in improved cryoprotection of human hematopoietic cells with reference to in vitro migration and adhesion properties".Transfusion 45, April 2005, 622-633 [Citation] [Link]

    Lalita M. Sasnoor, Vaijayanti P. Kale andLalita S. Limaye.
    "Supplementation of conventional freezing medium with a combination of catalase and trehalose results in better protection of surface molecules and functionality of haematopoietic cells." Journal of haematotherapy and stem cell research. 2003,12(5) 553-64 [Citation] [Link]

    L.S. Limayeand V.P. Kale,
    "Cryopreservation of human haematopoietic cells with membrane stabilizers and bioantioxidants as additives in the conventional freezingmedium", Journal of haematotherapy and stem cell research, Vol .10,Issue 5, Oct 2001 709-718[Citation] [Link]

    V.P.Kale and Limaye L.S.
    Stimulation of adult bone marrow by factors secreted by fetal liver haematopoietic cells: In Vitro evaluation using semi-solid clonal assay system. Stem Cells (1999)17: 107:116. [Citation] [Link]

    Limaye L.S.
    Bone Marrow Cryopreservation: Improved recovery due to Bioantioxidant additives in the freezing solution. Stem Cells 1997; 15, 353-358. [Citation][Link]

    Limaye L.S.,Bhopale M.K., Sengupta S.P., Renapurkar D.M. & Sharma K.D.
    Studies on modes of transmission of angiostrongylus cantonesis infection to experimental albino rats. Indian J. Helminthology 1992; 42 (2), 37-40.

    Limaye L.S., Pradhan V.R., Bhopale M.K., Renapurkar D.M. & Sharma K.D.
    Angiostrongylus cantonesis: Intermediate, Pratenic & definitive host studies in Greater Bombay Area. Helminthologia 1988; 25 (1), 31-39.

    Limaye L.S., Bhopale M.K., Pradhan V.R., Renapurkar D.M. & Sharma K.D.
    Serological investigations of a suspected case of human angiostrongyliasis. Indian J. of Pathology and Microbiology 1988; 31 (3), 248-250.

    Bhopale M.K.,Limaye L.S., Pradhan V.R., Renapurkar D.M. & Sharma K.D.
    Studies on suspected clinical and experimental Angiostrongyliasis: Serological responses. J. Hygiene, Epidemiology, Microbiology and Immunology 1985; 29(3), 28-288.[Citation] [Link]

    Limaye L.S.,Bhopale M.K., Renapurkar D.M. & Sharma K.D.
    The distribution of Angiostrongylus cantonensis (Chen) in the central nervous system of Laboratory rats. Folio Parasitologia (Praha) 1983; 30, 81-284. [Citation] [Link]

    Renapurkar D.M., Bhopale M.K., Limaye L.S.,& Sharma K.D.
    Prevalence of Angiostrongylus cantonensis infection in commensal rats in Bombay. J. Helmintho. 1982; 56, 345-349. [Citation] [Link]

    Conferences and Talks

    Invited lectures in various National Institutes, Colleges and Workshops as well as on National Radio/TV on topics like 'Stem Cell Banking', 'Applications of FACS', 'Low temperature biology of hematopoietic stem cells'.

    Presented papers in National and International conferences on hematopoietic stem cells.

    Memberships and Awards

    Life member of

    Indian society of cell biology

    Biotechnology society of India

    Indian women scientists association

    Indian association of Microbiologists of India

    Lab Members

    Technical Staff

    Mrs. Nikhat Firdaus Qayyum Khan, Technical Officer A

    Ph.d Students

    Kedar Limbkar, Ph.D. Student - ICMR SRF
    Project: Effect of Oral feeding of Nutraceuticals on hematopoiesis and thrombopoiesis in mice.

    Ankita Dhenge, Ph.D. Student - UGC SRF
    Project: Identification of cellular and molecular events during platelets generation.

    Sophia Fernandes, Ph.D. Student - UGC SRF
    Project: Studies on generation of induced pluripotent stem cells from umbilical cord tissue derived adult stem cells.

    Prajakta Shinde, Ph.D Student-DBT SRF
    Project: In depth study on generation of Dendritic cells from hematopoietic stem cells.

    Rutuja Kuhikar, Ph.D. Student- DBT SRF
    Project: Studies on differentiation of hematopoietic stem cells towards red and white blood cell lineages.

    Alumni/Ph.d Completed Students

    Dr. Lalita Sasnoor,
    Research Scientist
    VAMC-University of California, San Francisco (UCSF)

    Dr. ShreeKumar Balan,
    Post doc,C/o Dr. Marc DALOD, Centre D & No.39,Immunologie, Marseille Luminy Parc Scienctifique et, Technologique de Luminy, Case 906, 13288 Marseille Cedex 9

    Dr. Sangeetha V.M,
    Assistant Research Scientist, Room 901S, Dept. of Basic Science & Craniofacial Biology, New York University College of Dentistry, 345 East 24th street, New York 10010

    Dr. Namrata Shabrani,
    Designation-(Ex) Postdoctoral associate
    Address-Department of Pharmacology University of Illinois College of Medicine 835 S. Wolcott Ave., E403 (5080 CoMRB) Chicago, IL 60612,Tel: 312-355-5935.Fax: 312-996-1225

    Jeetendra Kumar,
    Postdoctoral scholar
    University of California San Diego

    Manasi Talwadekar,
    Post-doctoral fellow
    Tata Institute of Fundamental Research (TIFR), Colaba, Mumbai

    Darshana Kadekar
    Post-doctoral fellow
    Denmark Technical university (DTU) Copenhagen,Denmark