PMC:2063610 / 26560-34057 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/2063610","sourcedb":"PMC","sourceid":"2063610","source_url":"http://www.ncbi.nlm.nih.gov/pmc/2063610","text":"hESCs indirectly enhance and rejuvenate the regeneration of skeletal muscle\nWhile hESC properties were inhibited by aged differentiated muscle, the myogenic potential of aged satellite cells seemed to be enhanced by co-cultures with hESCs (Fig. 4A). Therefore, we further explored the enhancing and rejuvenating effects of hESCs on myogenic potential in vitro and in vivo, using human mesenchymal stem cells (hMSCs) as a negative control. First, we examined the effects of hESCs on myotube generation by co-culture with primary myoblasts freshly derived from activated-by-injury satellite cells (Conboy et al., 2003). As shown in Fig. 5A (Mb + hESC), primary myoblasts underwent very rapid and robust nascent myotube formation, when co-cultured with hESCs for 48 h in myoblast differentiation medium. Namely, remarkably large fused myotubes containing approximately 50–70 nuclei formed around hESCs colonies (Fig. 5A). In contrast, when co-cultured with hMSCs, myotube formation was no greater than in myoblast cultures alone (Fig. 5A, Mb + hMSC and Mb alone). Encouraged by these data, we analyzed the myogenic potential of young and old satellite cells co-cultured with hESCs for 48 h. As shown in Fig. 5B, hESCs conferred a much-enhanced myogenic capacity on both young and, importantly, old myofiber-associated satellite cells (rapid formation of desmin+ myogenic cells, many of which formed de novo multinucleated myotubes). Control co-cultures of these satellite cells with hMSCs displayed no enhanced myogenicity. In summary, while the myogenic potential (production of desmin+ fusion-competent cells) was more pronounced in young vs. old myofiber-associated satellite cells under all experimental conditions, a finding that is consistent with previous data (Conboy et al., 2003), a clear increase in myogenic potential of old satellite cells was noted in co-cultures with hESCs, as compared to control cultures devoid of hESCs (Fig. 4A,B).\nInterestingly, in addition to the rejuvenating effects of direct co-cultures shown in Fig. 5, soluble factors present in hESC-conditioned culture supernatants were also able to enhance myogenesis of aged satellite cells (Supplementary Fig. S5). Thus, in agreement with the notion that an established embryonic microniche antagonizes the inhibitory effects of the aged environment on stem cell responses (Fig. 3), the hESC-produced factors enhanced myogenic capacity of even old mouse satellite cells.\nEstablishing that hESC-produced factors enhance adult myogenesis and rejuvenate the regenerative capacity of even aged satellite cells in vitro prompted us to examine whether the regeneration of old injured muscle will be improved by hESC transplantation in vivo. Additionally, based on the data shown above, we speculated that even if the host's repair capacity is improved, hESCs themselves will not be efficiently maintained or expanded in the context of old systemic and local organ environments, and will not directly contribute to the repair of aged skeletal muscle. To test these hypotheses, we injected 5 × 105 hESCs or control hMSCs into the tibialis anterior (TA) and gastrocnemius muscles of young and old mice at 24 h after cardiotoxin-induced injury, when activation/proliferation of endogenous satellite cells normally begins (Conboy et al., 2003, 2005; Wagers \u0026 Conboy, 2005). To avoid immune response against hESC antigens, mice were immunosuppressed using FK506 (Ito \u0026 Tanaka, 1997; Dumont, 2000). Muscle was isolated 5 days post-injury, when nascent differentiated myofibers normally replace the damaged tissue (Conboy et al., 2003), and 10 µm cryosections were analyzed for the success in tissue repair using hematoxylin and eosin (H\u0026E) histochemistry and eMyHC immunodetection. H\u0026E analysis reveals newly formed myofibers, based on their smaller size and centrally located nuclei. Additionally, de novo myofibers in the damaged area appear positive for eMyHC, while undamaged myofibers remain negative. As shown in Fig. 6A and quantified in 6B, injection of hESCs significantly enhanced regeneration of skeletal muscle. Remarkably, this positive embryonic effect was especially pronounced in old tissue.\nFig. 6 Skeletal muscle regeneration following hESC transplantation is a balance between the inhibitory influence of aged niches and the rejuvenating effects of hESCs. Young and old tibialis anterior and gastrocnemius muscles were injured by cardiotoxin injection. hESCs or hMSCs were transplanted at the site of injury and were analyzed by cryosectioning at Day 5 after injury (as described in Experimental procedures). (A) Newly regenerated myofibers were detected using eMyHC-specific antibody (green) and staining with H\u0026E. In H\u0026E staining, newly regenerated areas contain smaller, immature myofibers with centrally located nuclei. Uninjured myofibers are much larger, by comparison, with peripherally restricted nuclei. Poorly regenerated areas lack new myofibers and contain areas of fibrosis and inflammation. eMyHC immunodetection is specific for regenerating areas of muscle only. Both assays showed dramatic enhancement of muscle regeneration in ‘old + hESC’ vs. ‘old + hMSC’. Regeneration improvement was also seen in young + hESC, as compared to young + hMSC. (B) Quantification of muscle regeneration was performed by analyzing the density of newly formed myofibers per mm2 of injury site, which is the volume that typically covers the whole injured area. Multiple, 10 µm H\u0026E sections were examined through the entire volume of injury in multiple, independently injured muscles. n = 20; * indicates P \u003c 0.001 (‘old + hMSC’ compared to young + hMSC and ‘old + hMSC’ compared to ‘old + hESC’. (C) H\u0026E and immunofluoresence staining for Oct4, and a human-specific antibody to NuMA, revealed the failure of hESCs to expand or persist in old, but the presence of hESCs in young muscle at 5 days post-transplantation. Hoechst (blue) labels nuclei. Importantly, such enhanced and rejuvenated muscle repair stems from an indirect induction, as hESCs themselves (or control hMSCs) did not physically contribute to the mouse myofibers, as judged by near absence (less than 0.1%) of human-specific NuMA+ nuclei in de novo desmin+ myofibers, analyzed through multiple injury sites. An example of one regenerated myofiber from young muscle injected with hESCs, with NuMA+ nucleus in a field of NuMA−/desmin+ mouse myofibers, is shown in Supplementary Fig. S6. No such NuMA+/desmin+ myofibers were detected in aged regenerated muscle (not shown).\nIn agreement with the in vitro data, establishing that aged systemic and local niches inhibit hESC proliferation and Oct4 expression (Figs 2 and 4 and Supplementary Fig. S2), hESCs failed to expand or even persist in old muscle, as judged by the absence of NuMA+/Oct4+ hESC-derived cells in the aged tissue. In contrast, colonies of Numa+/Oct4+ hESC-derived cells that did not undergo myogenic differentiation were easily detected in young regenerating muscle (Fig. 6C). This finding validates several technical aspects of these experiments, and confirms the contrasting effects of young and old systemic and local organ niches on hESC self-renewal.\nThese data further confirm and extrapolate our findings and demonstrate that when exposed to both aged systemic and local organ niches, hESCs fail to persist and do not contribute to tissue repair directly. At the same time, these embryonic cells indirectly but significantly improve the repair of aged injured muscle in vivo.","divisions":[{"label":"Title","span":{"begin":0,"end":75}},{"label":"Figure caption","span":{"begin":4173,"end":5929}}],"tracks":[{"project":"2_test","denotations":[{"id":"17381551-14645852-1491348","span":{"begin":611,"end":615},"obj":"14645852"},{"id":"17381551-14645852-1491349","span":{"begin":1781,"end":1785},"obj":"14645852"},{"id":"17381551-14645852-1491350","span":{"begin":3305,"end":3309},"obj":"14645852"},{"id":"17381551-3909880-1491351","span":{"begin":3311,"end":3315},"obj":"3909880"},{"id":"17381551-16143100-1491352","span":{"begin":3334,"end":3338},"obj":"16143100"},{"id":"17381551-9433807-1491353","span":{"begin":3443,"end":3447},"obj":"9433807"},{"id":"17381551-10702636-1491354","span":{"begin":3457,"end":3461},"obj":"10702636"},{"id":"17381551-14645852-1491355","span":{"begin":3594,"end":3598},"obj":"14645852"}],"attributes":[{"subj":"17381551-14645852-1491348","pred":"source","obj":"2_test"},{"subj":"17381551-14645852-1491349","pred":"source","obj":"2_test"},{"subj":"17381551-14645852-1491350","pred":"source","obj":"2_test"},{"subj":"17381551-3909880-1491351","pred":"source","obj":"2_test"},{"subj":"17381551-16143100-1491352","pred":"source","obj":"2_test"},{"subj":"17381551-9433807-1491353","pred":"source","obj":"2_test"},{"subj":"17381551-10702636-1491354","pred":"source","obj":"2_test"},{"subj":"17381551-14645852-1491355","pred":"source","obj":"2_test"}]},{"project":"CellFinder","denotations":[{"id":"T347","span":{"begin":3085,"end":3090},"obj":"CellType"},{"id":"T348","span":{"begin":1359,"end":1373},"obj":"CellType"},{"id":"T349","span":{"begin":3068,"end":3073},"obj":"CellType"},{"id":"T350","span":{"begin":5940,"end":5941},"obj":"GeneProtein"},{"id":"T351","span":{"begin":1459,"end":1474},"obj":"CellType"},{"id":"T352","span":{"begin":1543,"end":1551},"obj":"Anatomy"},{"id":"T353","span":{"begin":1419,"end":1427},"obj":"CellType"},{"id":"T354","span":{"begin":528,"end":537},"obj":"CellType"},{"id":"T355","span":{"begin":1480,"end":1485},"obj":"CellType"},{"id":"T356","span":{"begin":1666,"end":1681},"obj":"CellType"},{"id":"T357","span":{"begin":1808,"end":1816},"obj":"CellType"},{"id":"T358","span":{"begin":1577,"end":1583},"obj":"GeneProtein"},{"id":"T359","span":{"begin":473,"end":478},"obj":"CellType"},{"id":"T360","span":{"begin":6341,"end":6345},"obj":"GeneProtein"},{"id":"T361","span":{"begin":6329,"end":6334},"obj":"CellType"},{"id":"T362","span":{"begin":1315,"end":1330},"obj":"CellType"},{"id":"T363","span":{"begin":6375,"end":6381},"obj":"GeneProtein"},{"id":"T364","span":{"begin":1209,"end":1214},"obj":"CellType"},{"id":"T365","span":{"begin":1241,"end":1249},"obj":"Anatomy"},{"id":"T366","span":{"begin":3100,"end":3117},"obj":"Anatomy"},{"id":"T367","span":{"begin":6347,"end":6354},"obj":"CellComponent"},{"id":"T368","span":{"begin":2432,"end":2447},"obj":"CellType"},{"id":"T369","span":{"begin":1103,"end":1111},"obj":"Anatomy"},{"id":"T370","span":{"begin":6383,"end":6388},"obj":"Species"},{"id":"T371","span":{"begin":2152,"end":2167},"obj":"CellType"},{"id":"T372","span":{"begin":6854,"end":6858},"obj":"GeneProtein"},{"id":"T373","span":{"begin":6443,"end":6447},"obj":"GeneProtein"},{"id":"T374","span":{"begin":6784,"end":6788},"obj":"GeneProtein"},{"id":"T375","span":{"begin":1139,"end":1154},"obj":"CellType"},{"id":"T376","span":{"begin":6449,"end":6455},"obj":"GeneProtein"},{"id":"T377","span":{"begin":2248,"end":2257},"obj":"Anatomy"},{"id":"T378","span":{"begin":1172,"end":1177},"obj":"CellType"},{"id":"T379","span":{"begin":2568,"end":2583},"obj":"CellType"},{"id":"T380","span":{"begin":1880,"end":1885},"obj":"CellType"},{"id":"T381","span":{"begin":6053,"end":6058},"obj":"CellType"},{"id":"T382","span":{"begin":6103,"end":6112},"obj":"CellType"},{"id":"T383","span":{"begin":6097,"end":6112},"obj":"CellType"},{"id":"T384","span":{"begin":6906,"end":6914},"obj":"Anatomy"},{"id":"T385","span":{"begin":1050,"end":1052},"obj":"CellType"},{"id":"T386","span":{"begin":6199,"end":6205},"obj":"GeneProtein"},{"id":"T387","span":{"begin":6175,"end":6179},"obj":"GeneProtein"},{"id":"T388","span":{"begin":6181,"end":6187},"obj":"CellComponent"},{"id":"T389","span":{"begin":6207,"end":6216},"obj":"CellType"},{"id":"T390","span":{"begin":3005,"end":3013},"obj":"Anatomy"},{"id":"T391","span":{"begin":4115,"end":4124},"obj":"Anatomy"},{"id":"T392","span":{"begin":3257,"end":3272},"obj":"CellType"},{"id":"T393","span":{"begin":3842,"end":3848},"obj":"CellComponent"},{"id":"T394","span":{"begin":3119,"end":3121},"obj":"Anatomy"},{"id":"T395","span":{"begin":6024,"end":6029},"obj":"CellType"},{"id":"T396","span":{"begin":3872,"end":3881},"obj":"CellType"},{"id":"T397","span":{"begin":60,"end":68},"obj":"Anatomy"},{"id":"T398","span":{"begin":3532,"end":3541},"obj":"CellType"},{"id":"T399","span":{"begin":3464,"end":3470},"obj":"Anatomy"},{"id":"T400","span":{"begin":60,"end":75},"obj":"Anatomy"},{"id":"T401","span":{"begin":3127,"end":3148},"obj":"Anatomy"},{"id":"T402","span":{"begin":893,"end":898},"obj":"CellType"},{"id":"T403","span":{"begin":954,"end":959},"obj":"CellType"},{"id":"T404","span":{"begin":6097,"end":6102},"obj":"Species"},{"id":"T405","span":{"begin":4072,"end":4087},"obj":"Anatomy"},{"id":"T406","span":{"begin":6389,"end":6398},"obj":"CellType"},{"id":"T407","span":{"begin":0,"end":5},"obj":"CellType"},{"id":"T408","span":{"begin":4072,"end":4080},"obj":"Anatomy"},{"id":"T409","span":{"begin":397,"end":407},"obj":"CellType"},{"id":"T410","span":{"begin":4027,"end":4032},"obj":"CellType"},{"id":"T411","span":{"begin":385,"end":396},"obj":"Anatomy"},{"id":"T412","span":{"begin":3922,"end":3927},"obj":"GeneProtein"},{"id":"T413","span":{"begin":3945,"end":3954},"obj":"CellType"},{"id":"T414","span":{"begin":323,"end":328},"obj":"CellType"},{"id":"T415","span":{"begin":332,"end":340},"obj":"Anatomy"},{"id":"T416","span":{"begin":1036,"end":1038},"obj":"CellType"},{"id":"T417","span":{"begin":3005,"end":3020},"obj":"Anatomy"},{"id":"T418","span":{"begin":2825,"end":2830},"obj":"CellType"},{"id":"T419","span":{"begin":1404,"end":1418},"obj":"CellComponent"},{"id":"T420","span":{"begin":1351,"end":1357},"obj":"GeneProtein"},{"id":"T421","span":{"begin":750,"end":755},"obj":"CellType"},{"id":"T422","span":{"begin":6638,"end":6642},"obj":"GeneProtein"},{"id":"T423","span":{"begin":6860,"end":6864},"obj":"GeneProtein"},{"id":"T424","span":{"begin":659,"end":668},"obj":"CellType"},{"id":"T425","span":{"begin":6696,"end":6701},"obj":"CellType"},{"id":"T426","span":{"begin":148,"end":156},"obj":"Anatomy"},{"id":"T427","span":{"begin":579,"end":594},"obj":"CellType"},{"id":"T428","span":{"begin":175,"end":190},"obj":"CellType"},{"id":"T429","span":{"begin":639,"end":641},"obj":"CellType"},{"id":"T430","span":{"begin":701,"end":716},"obj":"CellType"},{"id":"T431","span":{"begin":379,"end":407},"obj":"CellType"},{"id":"T432","span":{"begin":233,"end":238},"obj":"CellType"},{"id":"T433","span":{"begin":409,"end":414},"obj":"CellType"},{"id":"T434","span":{"begin":6778,"end":6782},"obj":"GeneProtein"},{"id":"T435","span":{"begin":6443,"end":6465},"obj":"Anatomy"},{"id":"T436","span":{"begin":3389,"end":3393},"obj":"Species"},{"id":"T437","span":{"begin":6389,"end":6397},"obj":"Anatomy"},{"id":"T438","span":{"begin":3166,"end":3170},"obj":"Species"},{"id":"T439","span":{"begin":3781,"end":3789},"obj":"Anatomy"},{"id":"T440","span":{"begin":3532,"end":3540},"obj":"Anatomy"},{"id":"T441","span":{"begin":768,"end":775},"obj":"CellType"},{"id":"T442","span":{"begin":385,"end":406},"obj":"CellType"},{"id":"T443","span":{"begin":6288,"end":6296},"obj":"Anatomy"},{"id":"T444","span":{"begin":3872,"end":3880},"obj":"Anatomy"},{"id":"T445","span":{"begin":3945,"end":3953},"obj":"Anatomy"},{"id":"T446","span":{"begin":7402,"end":7417},"obj":"CellType"},{"id":"T447","span":{"begin":7307,"end":7312},"obj":"CellType"},{"id":"T448","span":{"begin":6457,"end":6466},"obj":"CellType"},{"id":"T449","span":{"begin":2426,"end":2431},"obj":"Species"},{"id":"T450","span":{"begin":3781,"end":3790},"obj":"CellType"},{"id":"T451","span":{"begin":1929,"end":1934},"obj":"CellType"},{"id":"T452","span":{"begin":1834,"end":1849},"obj":"CellType"},{"id":"T453","span":{"begin":6369,"end":6373},"obj":"GeneProtein"},{"id":"T454","span":{"begin":6103,"end":6111},"obj":"Anatomy"},{"id":"T455","span":{"begin":7402,"end":7411},"obj":"Anatomy"},{"id":"T456","span":{"begin":2396,"end":2404},"obj":"CellType"},{"id":"T457","span":{"begin":6199,"end":6215},"obj":"Anatomy"},{"id":"T458","span":{"begin":3724,"end":3729},"obj":"GeneProtein"},{"id":"T459","span":{"begin":2248,"end":2268},"obj":"Anatomy"},{"id":"T460","span":{"begin":1646,"end":1654},"obj":"Anatomy"},{"id":"T461","span":{"begin":832,"end":840},"obj":"CellType"},{"id":"T462","span":{"begin":872,"end":878},"obj":"CellComponent"},{"id":"T463","span":{"begin":826,"end":840},"obj":"CellType"},{"id":"T464","span":{"begin":1404,"end":1426},"obj":"Anatomy"},{"id":"T465","span":{"begin":1295,"end":1303},"obj":"Anatomy"},{"id":"T805","span":{"begin":4768,"end":4777},"obj":"CellType"},{"id":"T806","span":{"begin":4990,"end":4995},"obj":"GeneProtein"},{"id":"T807","span":{"begin":4932,"end":4941},"obj":"CellType"},{"id":"T808","span":{"begin":4646,"end":4651},"obj":"GeneProtein"},{"id":"T809","span":{"begin":4819,"end":4828},"obj":"CellType"},{"id":"T810","span":{"begin":4890,"end":4896},"obj":"CellComponent"},{"id":"T811","span":{"begin":4801,"end":4807},"obj":"CellComponent"},{"id":"T812","span":{"begin":5342,"end":5350},"obj":"Anatomy"},{"id":"T813","span":{"begin":4768,"end":4776},"obj":"Anatomy"},{"id":"T814","span":{"begin":4932,"end":4940},"obj":"Anatomy"},{"id":"T815","span":{"begin":5556,"end":5563},"obj":"Anatomy"},{"id":"T816","span":{"begin":4334,"end":4339},"obj":"CellType"},{"id":"T817","span":{"begin":4181,"end":4196},"obj":"Anatomy"},{"id":"T818","span":{"begin":5920,"end":5926},"obj":"CellComponent"},{"id":"T819","span":{"begin":4355,"end":4372},"obj":"Anatomy"},{"id":"T820","span":{"begin":4181,"end":4189},"obj":"Anatomy"},{"id":"T821","span":{"begin":5342,"end":5351},"obj":"CellType"},{"id":"T822","span":{"begin":4819,"end":4827},"obj":"Anatomy"},{"id":"T823","span":{"begin":4616,"end":4624},"obj":"Anatomy"},{"id":"T824","span":{"begin":4377,"end":4390},"obj":"Anatomy"},{"id":"T825","span":{"begin":4616,"end":4625},"obj":"CellType"},{"id":"T826","span":{"begin":4391,"end":4398},"obj":"Anatomy"},{"id":"T827","span":{"begin":5759,"end":5763},"obj":"GeneProtein"},{"id":"T828","span":{"begin":5789,"end":5794},"obj":"CellType"},{"id":"T829","span":{"begin":5720,"end":5724},"obj":"GeneProtein"},{"id":"T830","span":{"begin":5844,"end":5849},"obj":"CellType"},{"id":"T831","span":{"begin":4447,"end":4452},"obj":"CellType"},{"id":"T832","span":{"begin":4438,"end":4443},"obj":"CellType"}],"attributes":[{"subj":"T347","pred":"source","obj":"CellFinder"},{"subj":"T348","pred":"source","obj":"CellFinder"},{"subj":"T349","pred":"source","obj":"CellFinder"},{"subj":"T350","pred":"source","obj":"CellFinder"},{"subj":"T351","pred":"source","obj":"CellFinder"},{"subj":"T352","pred":"source","obj":"CellFinder"},{"subj":"T353","pred":"source","obj":"CellFinder"},{"subj":"T354","pred":"source","obj":"CellFinder"},{"subj":"T355","pred":"source","obj":"CellFinder"},{"subj":"T356","pred":"source","obj":"CellFinder"},{"subj":"T357","pred":"source","obj":"CellFinder"},{"subj":"T358","pred":"source","obj":"CellFinder"},{"subj":"T359","pred":"source","obj":"CellFinder"},{"subj":"T360","pred":"source","obj":"CellFinder"},{"subj":"T361","pred":"source","obj":"CellFinder"},{"subj":"T362","pred":"source","obj":"CellFinder"},{"subj":"T363","pred":"source","obj":"CellFinder"},{"subj":"T364","pred":"source","obj":"CellFinder"},{"subj":"T365","pred":"source","obj":"CellFinder"},{"subj":"T366","pred":"source","obj":"CellFinder"},{"subj":"T367","pred":"source","obj":"CellFinder"},{"subj":"T368","pred":"source","obj":"CellFinder"},{"subj":"T369","pred":"source","obj":"CellFinder"},{"subj":"T370","pred":"source","obj":"CellFinder"},{"subj":"T371","pred":"source","obj":"CellFinder"},{"subj":"T372","pred":"source","obj":"CellFinder"},{"subj":"T373","pred":"source","obj":"CellFinder"},{"subj":"T374","pred":"source","obj":"CellFinder"},{"subj":"T375","pred":"source","obj":"CellFinder"},{"subj":"T376","pred":"source","obj":"CellFinder"},{"subj":"T377","pred":"source","obj":"CellFinder"},{"subj":"T378","pred":"source","obj":"CellFinder"},{"subj":"T379","pred":"source","obj":"CellFinder"},{"subj":"T380","pred":"source","obj":"CellFinder"},{"subj":"T381","pred":"source","obj":"CellFinder"},{"subj":"T382","pred":"source","obj":"CellFinder"},{"subj":"T383","pred":"source","obj":"CellFinder"},{"subj":"T384","pred":"source","obj":"CellFinder"},{"subj":"T385","pred":"source","obj":"CellFinder"},{"subj":"T386","pred":"source","obj":"CellFinder"},{"subj":"T387","pred":"source","obj":"CellFinder"},{"subj":"T388","pred":"source","obj":"CellFinder"},{"subj":"T389","pred":"source","obj":"CellFinder"},{"subj":"T390","pred":"source","obj":"CellFinder"},{"subj":"T391","pred":"source","obj":"CellFinder"},{"subj":"T392","pred":"source","obj":"CellFinder"},{"subj":"T393","pred":"source","obj":"CellFinder"},{"subj":"T394","pred":"source","obj":"CellFinder"},{"subj":"T395","pred":"source","obj":"CellFinder"},{"subj":"T396","pred":"source","obj":"CellFinder"},{"subj":"T397","pred":"source","obj":"CellFinder"},{"subj":"T398","pred":"source","obj":"CellFinder"},{"subj":"T399","pred":"source","obj":"CellFinder"},{"subj":"T400","pred":"source","obj":"CellFinder"},{"subj":"T401","pred":"source","obj":"CellFinder"},{"subj":"T402","pred":"source","obj":"CellFinder"},{"subj":"T403","pred":"source","obj":"CellFinder"},{"subj":"T404","pred":"source","obj":"CellFinder"},{"subj":"T405","pred":"source","obj":"CellFinder"},{"subj":"T406","pred":"source","obj":"CellFinder"},{"subj":"T407","pred":"source","obj":"CellFinder"},{"subj":"T408","pred":"source","obj":"CellFinder"},{"subj":"T409","pred":"source","obj":"CellFinder"},{"subj":"T410","pred":"source","obj":"CellFinder"},{"subj":"T411","pred":"source","obj":"CellFinder"},{"subj":"T412","pred":"source","obj":"CellFinder"},{"subj":"T413","pred":"source","obj":"CellFinder"},{"subj":"T414","pred":"source","obj":"CellFinder"},{"subj":"T415","pred":"source","obj":"CellFinder"},{"subj":"T416","pred":"source","obj":"CellFinder"},{"subj":"T417","pred":"source","obj":"CellFinder"},{"subj":"T418","pred":"source","obj":"CellFinder"},{"subj":"T419","pred":"source","obj":"CellFinder"},{"subj":"T420","pred":"source","obj":"CellFinder"},{"subj":"T421","pred":"source","obj":"CellFinder"},{"subj":"T422","pred":"source","obj":"CellFinder"},{"subj":"T423","pred":"source","obj":"CellFinder"},{"subj":"T424","pred":"source","obj":"CellFinder"},{"subj":"T425","pred":"source","obj":"CellFinder"},{"subj":"T426","pred":"source","obj":"CellFinder"},{"subj":"T427","pred":"source","obj":"CellFinder"},{"subj":"T428","pred":"source","obj":"CellFinder"},{"subj":"T429","pred":"source","obj":"CellFinder"},{"subj":"T430","pred":"source","obj":"CellFinder"},{"subj":"T431","pred":"source","obj":"CellFinder"},{"subj":"T432","pred":"source","obj":"CellFinder"},{"subj":"T433","pred":"source","obj":"CellFinder"},{"subj":"T434","pred":"source","obj":"CellFinder"},{"subj":"T435","pred":"source","obj":"CellFinder"},{"subj":"T436","pred":"source","obj":"CellFinder"},{"subj":"T437","pred":"source","obj":"CellFinder"},{"subj":"T438","pred":"source","obj":"CellFinder"},{"subj":"T439","pred":"source","obj":"CellFinder"},{"subj":"T440","pred":"source","obj":"CellFinder"},{"subj":"T441","pred":"source","obj":"CellFinder"},{"subj":"T442","pred":"source","obj":"CellFinder"},{"subj":"T443","pred":"source","obj":"CellFinder"},{"subj":"T444","pred":"source","obj":"CellFinder"},{"subj":"T445","pred":"source","obj":"CellFinder"},{"subj":"T446","pred":"source","obj":"CellFinder"},{"subj":"T447","pred":"source","obj":"CellFinder"},{"subj":"T448","pred":"source","obj":"CellFinder"},{"subj":"T449","pred":"source","obj":"CellFinder"},{"subj":"T450","pred":"source","obj":"CellFinder"},{"subj":"T451","pred":"source","obj":"CellFinder"},{"subj":"T452","pred":"source","obj":"CellFinder"},{"subj":"T453","pred":"source","obj":"CellFinder"},{"subj":"T454","pred":"source","obj":"CellFinder"},{"subj":"T455","pred":"source","obj":"CellFinder"},{"subj":"T456","pred":"source","obj":"CellFinder"},{"subj":"T457","pred":"source","obj":"CellFinder"},{"subj":"T458","pred":"source","obj":"CellFinder"},{"subj":"T459","pred":"source","obj":"CellFinder"},{"subj":"T460","pred":"source","obj":"CellFinder"},{"subj":"T461","pred":"source","obj":"CellFinder"},{"subj":"T462","pred":"source","obj":"CellFinder"},{"subj":"T463","pred":"source","obj":"CellFinder"},{"subj":"T464","pred":"source","obj":"CellFinder"},{"subj":"T465","pred":"source","obj":"CellFinder"},{"subj":"T805","pred":"source","obj":"CellFinder"},{"subj":"T806","pred":"source","obj":"CellFinder"},{"subj":"T807","pred":"source","obj":"CellFinder"},{"subj":"T808","pred":"source","obj":"CellFinder"},{"subj":"T809","pred":"source","obj":"CellFinder"},{"subj":"T810","pred":"source","obj":"CellFinder"},{"subj":"T811","pred":"source","obj":"CellFinder"},{"subj":"T812","pred":"source","obj":"CellFinder"},{"subj":"T813","pred":"source","obj":"CellFinder"},{"subj":"T814","pred":"source","obj":"CellFinder"},{"subj":"T815","pred":"source","obj":"CellFinder"},{"subj":"T816","pred":"source","obj":"CellFinder"},{"subj":"T817","pred":"source","obj":"CellFinder"},{"subj":"T818","pred":"source","obj":"CellFinder"},{"subj":"T819","pred":"source","obj":"CellFinder"},{"subj":"T820","pred":"source","obj":"CellFinder"},{"subj":"T821","pred":"source","obj":"CellFinder"},{"subj":"T822","pred":"source","obj":"CellFinder"},{"subj":"T823","pred":"source","obj":"CellFinder"},{"subj":"T824","pred":"source","obj":"CellFinder"},{"subj":"T825","pred":"source","obj":"CellFinder"},{"subj":"T826","pred":"source","obj":"CellFinder"},{"subj":"T827","pred":"source","obj":"CellFinder"},{"subj":"T828","pred":"source","obj":"CellFinder"},{"subj":"T829","pred":"source","obj":"CellFinder"},{"subj":"T830","pred":"source","obj":"CellFinder"},{"subj":"T831","pred":"source","obj":"CellFinder"},{"subj":"T832","pred":"source","obj":"CellFinder"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#ec93ca","default":true},{"id":"CellFinder","color":"#93e4ec"}]}]}}