@article{d'costa_rich_diekman_2020, title={Engineered Cartilage from Human Chondrocytes with Homozygous Knockout of Cell Cycle Inhibitor p21}, volume={26}, ISSN={["1937-335X"]}, DOI={10.1089/ten.tea.2019.0214}, abstractNote={Osteoarthritis (OA) is a highly prevalent disease with limited treatment options. The search for disease-modifying OA therapies would benefit from a more comprehensive knowledge of the genetic variants that contribute to chondrocyte dysfunction and the barriers to cartilage regeneration. One goal of this study was to establish a system for producing engineered cartilage tissue from genetically defined primary human chondrocytes through genome editing and single-cell expansion. This process was utilized to investigate the functional effect of biallelic knockout of the cell cycle inhibitor p21. The use of ribonucleoprotein (RNP) CRISPR/Cas9 complexes targeting two sites in the coding region of p21 resulted in a high frequency (16%) of colonies with homozygous p21 knockout. Chondrogenic pellet cultures from expanded chondrocytes with complete loss of p21 produced more glycosaminoglycans (GAG) and maintained a higher cell number. Single-cell-derived colonies retained the potential for robust matrix production after expansion, allowing for analysis of colony variability from the same population of targeted cells. The effect of enhanced cartilage matrix production in p21 knockout chondrocytes persisted when matrix production from individual colonies was analyzed. Chondrocytes had lower levels of p21 protein with further expansion, and the difference in GAG production with p21 knockout was strongest at early passages. These results support previous findings that implicate p21 as a barrier to cartilage matrix production and regenerative capacity. Furthermore, this work establishes the use of genome-edited human chondrocytes as a promising approach for engineered tissue models containing user-defined gene knockouts and other genetic variants for investigation of OA pathogenesis. This work provides two important advances to the field of tissue engineering. One is the demonstration that engineered cartilage tissue can be produced from genetically defined populations of primary human chondrocytes. While CRISPR/Cas-9 genome editing has been extensively used in cell lines that divide indefinitely, this work extends the technique to an engineered tissue model system to support investigation of genetic changes that affect cartilage production. A second contribution is the finding that chondrocytes with p21 knockout synthesized more cartilage matrix tissue than unedited controls. This supports the continued investigation of p21 as a potential barrier to effective cartilage regeneration.}, number={7-8}, journal={TISSUE ENGINEERING PART A}, author={D'Costa, Susan and Rich, Matthew J. and Diekman, Brian O.}, year={2020}, month={Apr}, pages={441–449} }
 @article{boyer_s. d'costa_edwards_milloway_susick_borst_thakur_campbell_crenshaw_polo_et al._2015, title={Early-life dietary spray-dried plasma influences immunological and intestinal injury responses to later-life Salmonella typhimurium challenge}, volume={113}, ISSN={["1475-2662"]}, DOI={10.1017/s000711451400422x}, abstractNote={Increasing evidence supports the concept that early-life environmental influences, including nutrition and stress, have an impact on long-term health outcomes and disease susceptibility. The objective of the present study was to determine whether dietary spray-dried plasma (SDP), fed during the first 2 weeks post-weaning (PW), influences subsequent immunological and intestinal injury responses toSalmonellatyphimuriumchallenge. A total of thirty-two piglets (age 16–17 d) were weaned onto nursery diets containing 0, 2·5 % SDP (fed for 7 d PW) or 5 % SDP (fed for 14 d PW), and were then fed control diets (without SDP), for the remainder of the experiment. At 34 d PW (age 50 d), pigs were challenged with 3 × 109colony-forming units ofS. typhimurium. A control group (non-challenged) that was fed 0 % SDP in the nursery was included. At 2 d post-challenge, the distal ileum was harvested for the measurement of inflammatory, histological and intestinal physiological parameters.S.typhimuriumchallenge induced elevated ileal histological scores, myeloperoxidase (MPO), IL-8 and TNF, and increased intestinal permeability (indicated by reduced transepithelial voltage (potential difference) and elevated 4 kDa fluorescein isothiocyanate dextran (FD4) flux rates). Compared withS.typhimurium-challenged controls (0 % SDP), pigs fed the 5 % SDP-14 d diet exhibited reduced ileal histological scores, MPO levels, IL-8 levels and FD4 flux rates. Pigs fed the 5 % SDP-14 d nursery diet exhibited increased levels of plasma and ileal TNF-α in response to the challenge, compared with the other treatments. These results indicate that inclusion of SDP in PW diets can have an influence on subsequent immunological and intestinal injury responses induced by later-lifeS.typhimuriumchallenge.}, number={5}, journal={BRITISH JOURNAL OF NUTRITION}, author={Boyer, P. E. and S. D'Costa and Edwards, L. L. and Milloway, M. and Susick, E. and Borst, L. B. and Thakur, S. and Campbell, J. M. and Crenshaw, J. D. and Polo, J. and et al.}, year={2015}, month={Mar}, pages={783–793} }
 @article{d'costa_borst_kim_2013, title={Bone marrow-derived cells participate in the formation of normal and neoplastic lung stroma}, volume={33}, number={3}, journal={Anticancer Research}, author={D'Costa, S. and Borst, L. and Kim, Y.}, year={2013}, pages={831–836} }
 @article{kim_d'costa_suter_kim_2013, title={Evaluation of a side population of canine lymphoma cells using Hoechst 33342 dye}, volume={14}, ISSN={["1976-555X"]}, DOI={10.4142/jvs.2013.14.4.481}, abstractNote={Cancer stem cell (CSC) research has increased exponentially to gain further insight into the mechanisms underlying both carcinogenesis and chemotherapy resistance. The present study was performed to explore the potential value of a side population (SP) assay for identifying and characterizing putative CSCs among canine lymphoma cells. Canine lymphoma cells from cell lines and clinical samples were subjected to the SP assay consisting of Hoechst 33342 staining and subsequent flow cytometric analysis. The SP assay revealed various amounts of a SP fraction among the canine lymphoma cells. The percentages of SP were not affected by inhibitors of membrane transporters, verapamil hydrochloride, or fumitremorgin C. Most of the canine lymphoma cells expressed high levels of Bmi-1 and membrane transporter proteins such as ABCG2 and phosphorylated (p)-glycoprotein. This investigation lays the groundwork for further studies of the biological behaviors and molecular characteristics of CSCs in cases of canine lymphoma.}, number={4}, journal={JOURNAL OF VETERINARY SCIENCE}, author={Kim, Myung-Chul and D'Costa, Susan and Suter, Steven and Kim, Yongbaek}, year={2013}, month={Dec}, pages={481–486} }
 @article{s. d'costa_yoon_kim_motsinger-reif_williams_kim_2012, title={Morphologic and Molecular Analysis of 39 Spontaneous Feline Pulmonary Carcinomas}, volume={49}, ISSN={["1544-2217"]}, url={http://dx.doi.org/10.1177/0300985811419529}, DOI={10.1177/0300985811419529}, abstractNote={The present study was performed to determine the morphologic change and selected molecular features of spontaneous lung tumors in cats examined at the North Carolina State University Veterinary Teaching Hospital. Thirty-nine primary lung carcinomas represented 0.69% of all feline cases admitted to the hospital. Most lung tumors were observed in aged cats ( P < .0001), and no sex predilection was found ( P < .4241). Persian cats with pulmonary carcinoma were overrepresented in the data set, at least 4 times more frequently than other breeds. The histologic tumor types included adenocarcinoma (64.1%), bronchioloalveolar carcinoma (20.5%), and adenosquamous carcinoma (15.4%). Metastasis was observed in about 80% of 39 cases, with decreasing order of intrapulmonary metastasis, intrathoracic carcinomatosis, regional lymph nodes, and distant organs, including digits. The size of the largest tumor mass was significantly associated with metastatic potential ( P < .001). Based on immunohistochemistry, more than 80% (20 of 24) of feline lung tumors were positively labeled with either surfactant protein A or thyroid transcription factor 1. Epidermal growth factor receptor mutant and p53 proteins were detected in approximately 20% (5 of 24) and 25% (6 of 24) of the feline lung tumor cases, respectively. Limited sequencing analysis of K-ras and p53 genes in 3 selected normal and neoplastic lung tissues did not reveal any alteration. Results indicate that primary lung carcinomas are rare but aggressive tumors in cats, thereby warranting further studies on molecular carcinogenesis.}, number={6}, journal={VETERINARY PATHOLOGY}, author={S. D'Costa and Yoon, B. -I. and Kim, D. -Y. and Motsinger-Reif, A. A. and Williams, M. and Kim, Y.}, year={2012}, month={Nov}, pages={971–978} }
 @article{kai_d'costa_yoon_brody_sills_kim_2010, title={Characterization of side population cells in human malignant mesothelioma cell lines}, volume={70}, ISSN={["1872-8332"]}, DOI={10.1016/j.lungcan.2010.04.020}, abstractNote={Side population (SP) assay composed of Hoechst 33342 staining and subsequent flow cytometric analysis has been widely utilized for characterizing putative cancer stem cells (CSCs) in various human malignancies. The present study was designed to evaluate the SP assay as a research tool for mesothelial CSCs. A distinct fraction of SP cells was identified in various human malignant mesothelioma (HMM) cell lines, ranging from 0.05 to 1.32%. The sorted mesothelial SP cells exhibited enhanced proliferation potentials and higher expression of stem-cell genes, compared to non-SP (NSP) cells. Cisplatin treatment increased percentage of SP cells in the HMM cell lines. However, tumorigenic potential of SP cells in immunodeficient mice was similar to that of the NSP cells. These data indicated that SP assay may not be appropriate for enriching putative CSCs in HMM cell lines, and thus warrants the development of a novel tool for mesothelial CSC study.}, number={2}, journal={LUNG CANCER}, author={Kai, Kiyonori and D'Costa, Susan and Yoon, Byung-Il and Brody, Arnold R. and Sills, Robert C. and Kim, Yongbaek}, year={2010}, month={Nov}, pages={146–151} }
 @article{kai_d'costa_sills_kim_2009, title={Inhibition of the insulin-like growth factor 1 receptor pathway enhances the antitumor effect of cisplatin in human malignant mesothelioma cell lines}, volume={278}, ISSN={["1872-7980"]}, DOI={10.1016/j.canlet.2008.12.023}, abstractNote={Human malignant mesothelioma (HMM) is a fatal tumor and is poorly responsive to current therapeutic regimens. The insulin-like growth factor 1 receptor (IGF-1R) pathway is activated in HMM cell lines and tissues. Treatment with AG1024, an inhibitor of the IGF-1R pathway, significantly decreased cell proliferation and attenuated the phosphorylation of Akt and p44/42. In addition, it significantly enhanced the cytotoxic effects of cisplatin in HMM cell lines. This study supports the conjecture that inhibition of the IGF-1R pathway may be a useful target for reducing toxicity and alleviating chemoresistance to traditional anticancer drugs in HMM patients.}, number={1}, journal={CANCER LETTERS}, author={Kai, Kiyonori and D'Costa, Susan and Sills, Robert C. and Kim, Yongbaek}, year={2009}, month={Jun}, pages={49–55} }
 @article{song_s d'costa_pardue_petitte_2005, title={Production of germline chimeric chickens following the administration of a busulfan emulsion}, volume={70}, ISSN={["1098-2795"]}, url={http://europepmc.org/abstract/med/15685638}, DOI={10.1002/mrd.20218}, abstractNote={Busulfan (1,4‐butanediol dimethanesulfonate) was used to deplete endogenous germ cells for the enhanced production of chicken germline chimeras. Utilizing immunohistochemical identification of primordial gem cells (PGCs) in Stage 27 chicken embryos, two delivery formulations were compared relative to the degree of endogenous PGC depletion, a busulfan suspension (BS) and a solublized busulfan emulsion (SBE). Both busulfan treatments resulted in a significant reduction in PGCs when compared to controls. However, the SBE resulted in a more consistent and extensive depletion of PGCs than that observed with the BS treatment. Repopulation of SBE‐treated embryos with exogenous PGCs resulted in a threefold increase of PGCs in Stage 27 embryos. Subsequently, germline chimeras were produced by the transfer of male gonadal PGCs from Barred Plymouth Rock embryos into untreated and SBE‐treated White Leghorn embryos. Progeny testing of the presumptive chimeras with adult Barred Plymouth Rock chickens was performed to evaluate the efficiency of germline chimera production. The frequency of germline chimerism in SBE‐treated recipients increased fivefold when compared to untreated recipients. The number of donor‐derived offspring from the germline chimeras also increased eightfold following SBE‐treatment of the recipient embryos. These results demonstrated that the administration of a busulfan emulsion into the egg yolk of unincubated eggs improved the depletion of endogenous PGCs in the embryo and enhanced the efficiency of germline chimera production. Mol. Reprod. Dev. 70: 438–444, 2005. © 2005 Wiley‐Liss, Inc.}, number={4}, journal={MOLECULAR REPRODUCTION AND DEVELOPMENT}, author={Song, YH and S D'Costa and Pardue, SL and Petitte, JN}, year={2005}, month={Apr}, pages={438–444} }
 @misc{pardue_petitte_d'costa_song_2004, title={Methods for gamete production in birds}, volume={6,691,638}, number={2004 Feb. 17}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Pardue, S. and Petitte, J. and D'Costa, S. and Song, Y.-H.}, year={2004} }
 @misc{s d'costa_pardue_petitte_2001, title={Comparative development of avian primordial germ cells and production of germ line chimeras}, volume={12}, ISSN={["1470-2061"]}, DOI={10.3184/147020601783698477}, number={4}, journal={AVIAN AND POULTRY BIOLOGY REVIEWS}, author={S D'Costa and Pardue, SL and Petitte, JN}, year={2001}, pages={151–168} }
 @article{s d'costa_kulik_petitte_2000, title={Expression and purification of biologically active recombinant quail stem cell factor in E-coli}, volume={24}, ISSN={["1095-8355"]}, url={http://europepmc.org/abstract/med/10805965}, DOI={10.1006/cbir.1999.0500}, abstractNote={Abstract}, number={5}, journal={CELL BIOLOGY INTERNATIONAL}, author={S D'Costa and Kulik, MJ and Petitte, JN}, year={2000}, pages={311–317} }
 @misc{pardue_petitte_d'costa_2000, title={Methods for gamete production in birds}, volume={6,354,242}, number={2000 Mar 23}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Pardue, S. L. and Petitte, J. N. and D'Costa, S.}, year={2000} }
 @article{d'costa_petitte_1999, title={Characterization of stage-specific embryonic antigen-1 (SSEA-1) expression during early development of the turkey embryo}, volume={43}, number={4}, journal={International Journal of Developmental Biology}, author={D'Costa, S. and Petitte, J. N.}, year={1999}, pages={349–356} }