34 Íîâîñòè êëåòî÷íûõ òåõíîëîãèé 7.Kondziolka D., Wechsler L., Goldstein S. et al. Transplantation of cultured human neuronal cells for patients with stroke. Neurology 2000; 55: 565-9. 8.Nelson P.T., Kondziolka D., Wechsler L. et al. Clonal human (hNT) neuron grafts for stroke therapy: neuropathology in a patient 27 months after implantation. Am. J. Pathol. 2002; 160:1201-6. 9.Savitz S.I., Dinsmore J., Wu J. et al. Neurotransplantation of fetal porcine cells in patients with basal ganglia infarcts: a preliminary safety and feasibility study. Cerebrovasc. Dis. 2005; 20: 101-7. 10.Tateishi-Yuyama E., Matsubara H., Murohara T. et al. Therapeutic Angiogenesis using Cell Transplantation (TACT) Study Investigators. Therapeutic angiogenesis for patients with limb ischaemia by autologous transplantation of bone-marrow cells: a pilot study and a randomised controlled trial. Lancet 2002; 360: 427-35. 11.Strauer B.E., Brehm M., Zeus T. et al. Repair of infracted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans. Circ. 2002;106: 1913-8. 12.Chen J., Li Y., Wang L. et al. Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke 2001; 32: 1005-11. 13.Li Y., Chen J., Chen X.G. et al. Human marrow stromal cell therapy for stroke in rat: neurotrophins and functional recovery. Neurology 2002; 59: 514-23. 14.Zhao L.R., Duan W.M., Reyes M. et al. Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp. Neurol. 2002; 174: 11-20. 15.Chen J., Zhang Z.G., Li Y. et al. Intravenous administration of human bone marrow stromal cells induces angiogenesis in the ischemic boundary zone after stroke in rats. Circ. Res. 2003; 92: 692-9. 16.Massengale M., Wagers A.J., Vogel H., Weissman I.L. Hematopoietic cells maintain hematopoietic fates upon entering the brain. J. Exp. Med. 2005; 201: 1579-89. 17.Chen X., Li Y., Wang L. et al. Ischemic rat brain extracts induce human marrow stromal cell growth factor production. Neuropathology 2002; 22: 275-9. 18. Borlongan C.V., Hadman M., Sanberg C.D., Sanberg P.R. Central nervous system entry of peripherally injected umbilical cord blood cells is not required for neuroprotection in stroke. Stroke 2004; 35:2385-9. Ïîäãîòîâèë Ä.Ñ. Ñòàíêîâ ïî ìàòåðèàëàì Ann. Neurol. 2005; 57: 874-82 Èçó÷åíèå âûæèâàíèÿ è äèôôåðåíöèðîâêè àëëîãåííûõ ôåòàëüíûõ êëåòîê, òðàíñïëàíòèðîâàííûõ â ãîëîâíîé ìîçã ïàöèåíòàì ñ áîëåçíüþ Ïàðêèíñîíà - ðåçóëüòàòû èññëåäîâàíèÿ àóòîïñèéíîãî ìàòåðèàëà Ïåðâàÿ òðàíñïëàíòàöèÿ ôåòàëüíîé íåðâíîé òêàíè â ãîëîâíîé ìîçã ïàöèåíòàì ñ áîëåçíüþ Ïàðêèíñîíà (ÁÏ) áûëà âûïîëíåíà â êëèíèêå â 1987 ãîäó ãðóïïîé Lindvall O. â Lund University Hospital (Sweden) [1, 2]. C òåõ ïîð ñðàçó â íåñêîëüêèõ öåíòðàõ ìèðà áûëè íà÷àòû êëèíè÷åñêèå èñïûòàíèÿ ìåòîäà ñòåðåîòàêñè÷åñêîé íèãðîñòðèàëüíîé òðàíñïëàíòàöèè ôåòàëüíîé íåðâíîé òêàíè. Åâðîïåéñêàÿ ïðîãðàììà NECTAR (Network of European CNS Transplantation And Restoration), ïðèçâàííàÿ îöåíèòü êëèíè÷åñêóþ ýôôåêòèâíîñòü ìåòîäà, áûëà íà÷àòà â 1990 ãîäó è âêëþ÷àëà îêîëî 40 êëèíè÷åñêèõ ãðóïï.  ðåçóëüòàòå ýòèõ èñïûòàíèé áûëè ñäåëàíû âûâîäû, ÷òî ôåòàëüíûå àëëîãåííûå íèãðàëüíûå íåéðîíû ïðèæèâàþòñÿ áåç âûðàæåííîãî èììóííîãî îòâåòà, ðåèííåðâèðóþò îêðóæàþùèé ñòðèàòóì ãîëîâíîãî ìîçãà õîçÿèíà (÷åëîâåêà), âûçûâàþò îáðàçîâàíèå è âûáðîñ äîïàìèíà è îáåñïå÷èâàþò óëó÷øåíèå ìîòîðíîé ôóíêöèè [2, 3, 6]. Îäíàêî, â äâóõ äâîéíûõ-ñëåïûõ, ïëàöåáî-êîíòðîëèðóåìûõ (ïàöèåíòû áûëè ëîæíîîïåðèðîâàíû) èñïûòàíèÿõ, ñïîíñèðóåìûõ NIH â ÑØÀ, íå áûëî âûÿâëåíî çíà÷èìûõ ðàçëè÷èé â óëó÷øåíèè ìîòîðíîé ôóíêöèè ìåæäó ãðóïïàìè è íåîæèäàííî áûëè ïîëó÷åíû ïîáî÷íûå ýôôåêòû ó íåêîòîðûõ ïàöèåíòîâ [4, 5]. Ïåðâûå ìîðôîëîãè÷åñêèå èññëåäîâàíèÿ, ïîäòâåðæäàþùèå äîëãîâðåìåííîå âûæèâàíèå ïåðåñàæåííûõ êëåòîê è ÷àñòè÷íóþ ðåèííåðâàöèþ íèãðàñòðèàëüíîé îáëàñòè õîçÿèíà, áûëè âûïîëíåíû â 1995 ãîäó íà àóòîïñèéíîì ìàòåðèàëå ãîëîâíîãî ìîçãà ïàöèåíòîâ, ïîäâåðãøèõñÿ ïðîöåäóðå. Ýòè æå äàííûå áûëè ïîäòâåðæäåíû â áîëåå ñîâðåìåííûõ èññëåäîâàíèÿõ Olanow C.W. [5] è Freed C.R. [4]. Âñå ýòè èññëåäîâàíèÿ àíàëèçèðîâàëè ñóäüáó ôðàãìåíòîâ òêàíåâîãî òðàíñïëàíòàòà, íî íå ñóñïåíçèè íåéðîêëåòîê, ïîäâåðãøèõñÿ ïðåäâàðèòåëüíîé ýíçèìàòè÷åñêîé îáðàáîòêå è êóëüòèâèðîâàíèþ. Îäíàêî, èìåííî ïî ïðè÷èíå ïðèìåíåíèÿ ðàçëè÷íûõ âèäîâ òðàíñïëàíòàòîâ ðÿä èññëåäîâàòåëåé âèäèò ðàçíèöó, ïîëó÷åííóþ â êëèíè÷åñêèõ èññëåäîâàíèÿõ íåçàâèñèìûõ ãðóïï. Òàêèì îáðàçîì, èñïîëüçîâàíèå òêàíåâîãî èëè Êëåòî÷íàÿ òðàíñïëàíòîëîãèÿ è òêàíåâàÿ èíæåíåðèÿ ¹ 2, 2005 ñóñïåíçèîííîãî òðàíñïëàíòàòà îñòà¸òñÿ íåðåø¸ííûì âîïðîñîì êëèíè÷åñêîé íåéðîòðàíñïëàíòàöèè.  æóðíàëå Brain îïóáëèêîâàíû ðåçóëüòàòû ïàòîìîðôîëîãè÷åñêîãî èññëåäîâàíèÿ àóòîïñèéíîãî ìàòåðèàëà ãîëîâíîãî ìîçãà ïàöèåíòîâ ñ ÁÏ, ïîëó÷èâøèõ ñóñïåíçèîííûé íåéðîòðàíñïëàíòàò áîëåå 3-õ ëåò íàçàä. Ïðîäóêöèÿ äîôàìèíà ïî äàííûì F-18-DOPA ïîçèòðîííî-ýìèññèîííîé òîìîãðàôèè ãîëîâíîãî ìîçãà. Ñëåâà - äî ëå÷åíèÿ, ñïðàâà ÷åðåç 3 ãîäà ïîñëå òðàíñïëàíòàöèè. Èç Brain 2005; 128: 1498-510 Àíàëèçó ïîäâåðãëè òêàíü ãîëîâíîãî ìîçãà 2-õ ðåöèïèåíòîâ, ïîëó÷èâøèõ òðàíñïëàíòàöèè ôåòàëüíûõ êëåòîê è óìåðøèõ îò îñëîæíåíèé îñòðîãî èíôàðêòà ìèîêàðäà ÷åðåç 3 ãîäà 8 ìåñÿöåâ è 4 ãîäà 4 ìåñÿöà ïîñëå âìåøàòåëüñòâà. Âàæíî, ÷òî â ýòîì èññëåäîâàíèè ïåðåñàæèâàëè ñóñïåíçèþ êëåòîê, âûäåëåííóþ ýíçèìàòè÷åñêè (èç òêàíè ñðåäíåãî ìîçãà 6-9 íåäåëüíûõ ýìáðèîíîâ), ïðåäâàðèòåëüíî êóëüòèâèðîâàííûõ â ñðåäå ñ ôàêòîðàìè ðîñòà (GDNF). Òðàíñïëàíòàöèè âûïîëíÿëè ñòåðåîòàêñè÷åñêè áèëàòåðàëüíî â íåñêîëüêî òî÷åê ïî ðàçíûì òðàåêòîðèÿì â îáëàñòè ñòðèàòóìà (ïóòàìåí) è ÷¸ðíîé ñóáñòàíöèè. Ïåðâûé ïàöèåíò ïîëó÷èë 3,2 ìëí êëåòîê, âòîðîé - 4,8 ìëí (îáà - îò 3-õ ýìáðèîíîâ). Ïîñëå òðàíñïëàíòàöèè îáà ïàöèåíòà ïðîäåìîíñòðèðîâàëè êëèíè÷åñêîå óëó÷øåíèå è ïðèçíàêè ñèíòåçà äîïàìèíà â îáëàñòè òðàíñïëàíòàöèè. Àâòîðû ïîäñ÷èòàëè êîëè÷åñòâî äîïàìèí-íåéðîíîâ. Áîëüøàÿ ÷àñòü äîíîðñêèõ êëåòîê (40-60%) èìåëà ôåíîòèï 35 Human study äîïàìèí-íåéðîíîâ äâèãàòåëüíîé çîíû ñòðèàòóìà íîðìàëüíîãî ìîçãà ÷åëîâåêà (Calbidin-/Girk2+). Äîïàìèí-íåéðîíû òðàíñïëàíòàòà íàõîäèëè â êàæäîì êàíàëå ââåäåíèÿ êëåòîê è â âèäå ñêîïëåíèé - â ìåñòàõ ãðàôòèíãà. Îáùåå êîëè÷åñòâî òàêèõ íåéðîíîâ ñîñòàâëÿëî îêîëî 120-200 òûñÿ÷. Íåñìîòðÿ íà òî, ÷òî ïàöèåíòû ïðèíèìàëè èììóíîñóïðåññîðû (öèêëîñïîðèí À) â òå÷åíèå ïîëóãîäà ïîñëå òðàíñïëàíòàöèè, áûëè âûÿâëåíû ëèøü ïðèçíàêè íåáîëüøîãî ñêîïëåíèÿ (ïîÿâëåíèÿ òàê íàçûâàåìûõ «ðóáöîâ») CD45+ êëåòîê àêòèâèðîâàííîé ìèêðîãëèè (GFAP+) è åäèíè÷íûå ìàêðîôàãè. Ýòè äàííûå ïðîòèâîðå÷àò ìîðôîëîãè÷åñêèì äàííûì, ïîëó÷åííûì â èññëåäîâàíèè Freed C.R. [4], ãäå áûëà âûÿâëåíà âûðàæåííàÿ ìèêðîãëèàëüíàÿ ðåàêöèÿ è ïîâûøåííàÿ ýêñïðåññèÿ MHC-II (ãåíîâ ãèñòîñîâìåñòèìîñòè) ïðè òðàíñïëàíòàöèè ôðàãìåíòîâ òêàíè. Òàêèì îáðàçîì, â èññëåäîâàíèè áûëî ïîêàçàíî, ÷òî ôåòàëüíûå íåéðîíû, òðàíñïëàíòèðîâàííûå â ãîëîâíîé ìîçã ïàöèåíòîâ ñ áîëåçíüþ Ïàðêèíñîíà, äëèòåëüíî âûæèâàþò (áîëåå 3-4 ëåò). Êëåòêè ïðîäóöèðóþò òåðàïåâòè÷åñêèé óðîâåíü äîïàìèíà è ðåèííåðâèðóþò ïóòàìåí õîçÿèíà áåç ïîáî÷íûõ ýôôåêòîâ. Àâòîðû óêàçûâàþò, ÷òî ïîñëå òðàíñïëàíòàöèè â îáëàñòü ïóòàìåíà âûæèâàåìîñòü äîïàìèí+ êëåòîê ñîñòàâèëà 15-30%, à òðàíñïëàíòàòà îáëàñòè ÷¸ðíîé ñóáñòàíöèè 4-8%. Õîðîøèé ýôôåêò òðàíñïëàíòàöèè àâòîðû îáúÿñíÿþò ïðèìåíåíèåì ìåòîäà ïðåêóëüòèâèðîâàíèÿ ñóñïåíçèè íåéðîíîâ â ñðåäå ñ GDNF è ââåäåíèÿ èìåííî ñóñïåíçèè êëåòîê, ËÈÒÅÐÀÒÓÐÀ: 1. Lindvall O., Brundin P., Widner H. et al. Grafts of fetal dopamine neurons survive and improve motor function in Parkinson's disease. Science 1990; 247: 574-7. 2. Winkler C., Kirik D., Bjorklund A. Cell transplantation in Parkinson's disease: how can we make it work? Trends Neurosci. 2005; 28: 86-92. 3. Dunnett S.B., Bjorklund A., Lindvall O. Cell therapy in Parkinson's disease stop or go? Nat Rev. Neurosci. 2001; 2: 365-9. 4. Freed C.R., Greene P.E., Breeze R.E. et al. Transplantation of embryonic à íå ôðàãìåíòîâ òêàíè, êàê ýòî áûëî âûïîëíåíî â äâóõ ïðåäûäóùèõ èññëåäîâàíèÿõ. Êðîìå òîãî, â ýòîì èññëåäîâàíèè âïåðâûå áûëî ïðîàíàëèçèðîâàíî ñîñòîÿíèå òðàíñïëàíòàòà ïîñëå ãðàôòèíãà â îáëàñòü ÷¸ðíîé ñóáñòàíöèè. Îáùåå ÷èñëî âûæèâøèõ äîïàìèí+ íåéðîíîâ áûëî çíà÷èòåëüíî âûøå, ÷åì èõ ïðèìåðíîå êîëè÷åñòâî â òâ¸ðäûõ òðàíñïëàíòàòàõ, óêàçàííîå àâòîðàìè [4, 5]. Àâòîðû èññëåäîâàíèÿ óêàçûâàþò, ÷òî ïðèìåíåíèå ïîäîáíîãî ìåòîäà çàìåñòèòåëüíîé êëåòî÷íîé òðàíñïëàíòàöèè íå èìååò ïîáî÷íûõ ýôôåêòîâ è ìîæåò áûòü òåðàïåâòè÷åñêè âûãîäíûì äàæå ó ïàöèåíòîâ íà äàëåêî çàøåäøèõ ñòàäèÿõ çàáîëåâàíèÿ ñ àíàìíåçîì áîëåå 10 ëåò. Íàïðèìåð, ó îäíîãî ïàöèåíòà áûëî äîñòèãíóòî äîëãîâðåìåííîå âîññòàíîâëåíèå ñèíòåçà äîïàìèíà ñëåâà íà 65% è ñïðàâà - íà 88% îò íîðìàëüíîãî óðîâíÿ. Ïîñëå èñïûòàíèé ìåòîäà íåéðîòðàíñïëàíòàöèè äëÿ ëå÷åíèÿ áîëåçíè Ïàðêèíñîíà Olanow [5] è Freed [4], åãî êëèíè÷åñêàÿ ñóäüáà áûëà ïîñòàâëåíà ïîä ñîìíåíèå. Îäíàêî, òùàòåëüíûå ìîðôîëîãè÷åñêèå èññëåäîâàíèÿ ïîñòìîðòàëüíûõ òðàíñïëàíòàòîâ â ãîëîâíîì ìîçãå ðåöèïèåíòîâ ïîêàçûâàþò, ÷òî ìåòîä «ðàáîòàåò» è ìîæåò áûòü êëèíè÷åñêè âûãîäíûì äëÿ áîëüøèíñòâà ïàöèåíòîâ. Òàêèå èññëåäîâàíèÿ óêàçûâàþò êëèíèöèñòàì, êàêîé èìåííî ïðîòîêîë íåéðîòðàíñïëàíòàöèè îïòèìàëåí è ìîæåò ñëóæèòü ìåòîäîì âûáîðà. Äèñêóññèÿ î êëèíè÷åñêîé ñóäüáå è ïåðñïåêòèâàõ ìåòîäà ïðåäñòàâëåíà â íåäàâíåì îáçîðå [2]. dopamine neurons for severe Parkinson's disease. N. Engl. J. Med. 2001; 344: 710-9. 5. Olanow C.W., Goetz C.G., Kordower J.H. et al. A double-blind controlled trial of bilateral fetal nigral transplantation in Parkinson's disease. Ann. Neurol. 2003; 54: 403-14. 6. Kordower J.H., Freeman T.B., Snow B.J. et al. Neuropathological evidence of graft survival and striatal reinnervation after the transplantation of fetal mesencephalic tissue in a patient with Parkinson's disease. N. Engl. J. Med. 1995; 332: 1118-24. Ïîäãîòîâèë À.Â. Áåðñåíåâ ïî ìàòåðèàëàì Brain 2005; 128: 1498-510; doi:10.1093/brain/awh510 Òðàíñïëàíòàöèÿ êëåòîê êîñòíîãî ìîçãà ïàöèåíòàì ñ òðàâìîé ñïèííîãî ìîçãà - ðåçóëüòàòû ïèëîòíîãî êëèíè÷åñêîãî èññëåäîâàíèÿ â Þæíîé Êîðåå Òðàâìà ïîçâîíî÷íèêà, îñëîæíåííàÿ ïîâðåæäåíèåì ñïèííîãî ìîçãà â âèäå åãî êîìïðåññèè, ðàçìîçæåíèÿ, ÷àñòè÷íîãî èëè ïîëíîãî ðàçðûâà, ÿâëÿåòñÿ îäíîé èç àêòóàëüíûõ ìåäèêî-ñîöèàëüíûõ ïðîáëåì ñîâðåìåííîé ìåäèöèíû, ïîñêîëüêó âåä¸ò ê ãëóáîêîé èíâàëèäèçàöèè ïîñòðàäàâøèõ. Ïîñëåäñòâèÿ òðàâìû ñïèííîãî ìîçãà (ÒÑÌ) ÷àùå âñåãî ïðîÿâëÿþòñÿ âÿëûì èëè ñïàñòè÷åñêèì ïàðàëè÷îì, â ëó÷øåì ñëó÷àå - ïàðåçîì êîíå÷íîñòåé è äèñôóíêöèåé òàçîâûõ îðãàíîâ.  íàñòîÿùåå âðåìÿ íå ñóùåñòâóåò äåéñòâèòåëüíî ýôôåêòèâíûõ ìåòîäîâ ëå÷åíèÿ òðàâìàòè÷åñêîãî ïîâðåæäåíèÿ ñïèííîãî ìîçãà, îñîáåííî êîãäà ïîñëå òðàâìû ïðîøëè ìåñÿöû è ãîäû [1].  ïîñëåäíèå ãîäû áûëè èñïûòàíû ðàçëè÷íûå ìåòîäû êëåòî÷íîé íåéðîòðàíñïëàíòàöèè äëÿ êîððåêöèè ÒÑÌ â êëèíèêå [2]. Äëÿ ýòîãî ïðèìåíÿëè òðàíñïëàíòàöèþ ïîääåðæèâàþùèõ êëåòîê «îáîíÿòåëüíîãî ýïèòåëèÿ» [3], àêòèâèðîâàííûõ ìàêðîôàãîâ [4], ôåòàëüíûõ íåéðîíîâ [5] è êëåòîê êîñòíîãî ìîçãà [6].  ïðîøëîì ãîäó íà åæåãîäíîé êîíôåðåíöèè American Society for Neurotransplantation and Repair áûë ïðåäñòàâëåí ïåðâûé äîêëàä ïî ðåçóëüòàòàì àóòîëîãè÷íîé òðàíñïëàíòàöèè êëåòîê êîñòíîãî ìîçãà äëÿ ëå÷åíèÿ ÒÑÌ â Motol Hospital (Prague) [6]. Îò 10 äî 147 ìëí CD34+ êëåòîê êîñòíîãî ìîçãà ââîäèëè â ïîçâîíî÷íûå àðòåðèè 9-òè ïàöèåíòàì ñ ðàçëè÷íûìè óðîâíÿìè ïîâðåæäåíèÿ. Ó 6-òè èç 9-òè ïàöèåíòîâ áûëî îòìå÷åíî óëó÷øåíèå ÷åðåç 3-6 ìåñÿöåâ ïîñëå òðàíñïëàíòàöèè êëåòîê [2, 6].  íåäàâíåì íîìåðå æóðíàëà Tissue Engineering îïóáëèêîâàíû ðåçóëüòàòû êëèíè÷åñêîãî èññëåäîâàíèÿ ïî ýôôåêòèâíîñòè òðàíñïëàíòàöèè êëåòîê êîñòíîãî ìîçãà â îáëàñòü ïîâðåæäåíèÿ ñïèííîãî ìîçãà â ñî÷åòàíèè ñ òåðàïèåé ãðàíóëîöèòàðíî-ìàêðîôàãàëüíûì êîëîíèåñòèìóëèðóþùèì Êëåòî÷íàÿ òðàíñïëàíòîëîãèÿ è òêàíåâàÿ èíæåíåðèÿ ¹ 2, 2005