METHOD FOR AMPLIFYING CONE PHOTORECEPTORS OR ROD PHOTORECEPTORS USING DORSALIZATION SIGNAL TRANSMITTER OR VENTRALIZATION SIGNAL TRANSMITTER

1 . A method for increasing a proportion of a cone photoreceptor precursor and a cone photoreceptor in a photoreceptor precursor and a photoreceptor comprised in a retinal tissue, the method comprising a step of culturing a retinal tissue in an initial developmental stage to a stage where an emergence rate of a cone photoreceptor precursor reaches maximum in a medium comprising a dorsalization signal transmitter at a concentration sufficient to suppress expression of a ventral marker.

2 . The method according to claim 1 , wherein the cone photoreceptor precursor and the cone photoreceptor are CRX-positive and RXR-γ-positive, or CRX-positive and TRβ2-positive; and NRL-negative cells.

3 . The method according to claim 1 , wherein the ventral marker is ALDH1A3 and/or COUP-TF I.

4 . The method according to claim 1 , wherein the concentration of the dorsalization signal transmitter is such that it does not induce expression of a most dorsal marker.

5 . The method according to claim 1 , wherein the concentration of the dorsalization signal transmitter is such that it promotes expression of the dorsal marker.

6 . The method according to claim 1 , wherein the concentration of the dorsalization signal transmitter is such that it does not induce expression of the most dorsal marker and promotes expression of other dorsal markers.

7 . The method according to claim 5 , wherein the dorsal marker is CYP26A1 and/or CYP26C1.

8 . The method according to claim 4 , wherein the most dorsal marker is COUP-TF II.

9 . The method according to claim 5 , wherein the dorsal marker is ALDH1A1.

10 . The method according to claim 9 , wherein the concentration of the dorsalization signal transmitter is sufficient to induce expression of not less than 0.1% and not more than 30% of the expression level of ALDH1A1 promoted by 1.35 nM BMP4.

11 . The method according to claim 1 , wherein the retinal tissue in an initial developmental stage comprises (i) a ciliary marginal zone-like structure, or (ii) a cell that can differentiate into a photoreceptor and a ganglion cell.

12 . (canceled)

13 . The method according to claim 1 , wherein the retinal tissue in an initial developmental stage is derived from (i) a pluripotent stem cell or (ii) a neuroepithelial cell obtained from an adult tissue.

14 . (canceled)

15 . The method according to claim 1 , wherein the retinal tissue in an initial developmental stage comprises a PAX6-positive and RX-positive cell.

16 . The method according to claim 1 , wherein the retinal tissue in an initial developmental stage comprises a PAX6-positive, RX-positive and CHX10-positive cell.

17 . The method according to claim 1 , wherein the step of culturing in the presence of a dorsalization signal transmitter is continued for 4 days to 170 days.

18 . The method according to claim 17 , wherein the step of culturing in the presence of a dorsalization signal transmitter is continued until a period when a rod photoreceptor precursor emerges when cultured in the absence of a dorsalization signal transmitter.

19 . The method according to claim 1 , wherein the dorsalization signal transmitter is a BMP signal transduction pathway agonist or a Wnt signal transduction pathway agonist, or a SHH signal transduction pathway inhibitor which is capable of inducing a BMP signal corresponding to 0.01 nM-0.90 nM of BMP4.

20 . The method according to claim 1 , wherein the dorsalization signal transmitter is BMP4.

21 . The method according to claim 20 , wherein the concentration of BMP4 is 0.05 nM-0.45 nM.

22 . The method according to claim 1 , wherein the dorsalization signal transmitter is Cyclopamine-KAAD.

23 . The method according to claim 22 , wherein the concentration of Cyclopamine-KAAD is 0.01 μM-5 μM.

24 . (canceled)

25 . The method according to claim 1 , wherein the method is performed in a medium free of 9-cisretinoic acid.

26 . A retinal tissue comprising a photoreceptor precursor rich in a cone photoreceptor precursor and/or a photoreceptor rich in a cone photoreceptor, wherein the retinal tissue is obtained by the method according to claim 1 .

27 . A retinal tissue comprising a photoreceptor precursor rich in a cone photoreceptor precursor and/or a photoreceptor rich in a cone photoreceptor, and a ganglion cell, wherein the number of the cone photoreceptor precursor and cone photoreceptor is not less than 2 times, preferably not less than 4 times, the number of the rod photoreceptor precursor and rod photoreceptor, in the photoreceptor precursor and photoreceptor.

28 . The retinal tissue according to claim 27 , wherein the whole photoreceptor precursor and the whole photoreceptor comprises the cone photoreceptor precursor and cone photoreceptor in not less than 70%, preferably not less than 80%.

29 . A retinal tissue that is able to mature into the retinal tissue according to claim 27 by culturing.

30 . The retinal tissue according to claim 26 or 27 , wherein not less than 50% of the layer structure of the retinal tissue forms a continuous epithelial structure.

31 . The retinal tissue according to claim 30 , wherein the retinal tissue has a diameter in the major axis direction of not less than 0.6 mm.

32 . A pharmaceutical composition for transplantation to a retinal tissue of a retina disease patient in need of transplantation, comprising a retinal tissue section cut out from the retinal tissue according to claim 26 or 27 .

33 . The pharmaceutical composition according to claim 32 , wherein the retinal tissue requiring transplantation is a tissue of a region comprising Rod-free zone.

34 . The pharmaceutical composition according to claim 33 , wherein the region comprising the Rod-free zone has a macular-like structure.

35 . A method for increasing a proportion of a rod photoreceptor precursor and a rod photoreceptor in a photoreceptor precursor and a photoreceptor comprised in a retinal tissue, comprising a step of culturing a retinal tissue, in an initial developmental stage to a stage where an emergence rate of a cone photoreceptor precursor reaches maximum, for at least one day in the presence of a ventralization signal transmitter at a concentration sufficient to promote expression of a ventral marker.

36 . The method according to claim 35 , wherein the rod photoreceptor precursor and rod photoreceptor are NRL-positive and CRX-positive cells.

37 . The method according to claim 35 , wherein the ventral marker is ALDH1A3 and/or COUP-TF I.

38 . The method according to claim 35 , wherein the retinal tissue in an initial developmental stage comprises (i) a ciliary marginal zone-like structure, or (ii) a cell that can differentiate into photoreceptor or a ganglion cell.

39 . (canceled)

40 . The method according to claim 35 , wherein the retinal tissue in an initial developmental stage is derived from (i) a pluripotent stem cell, or (ii) a neuroepithelial cell obtained from an adult tissue.

41 . (canceled)

42 . The method according to claim 35 , wherein the retinal tissue in an initial developmental stage comprises a PAX6-positive and RX-positive cell.

43 . The method according to claim 42 , wherein the retinal tissue in an initial developmental stage comprises a PAX6-positive, RX-positive and CHX10-positive cell.

44 . The method according to claim 35 , wherein the step of culturing in the presence of a ventralization signal transmitter is continued for 4 days to 170 days.

45 . The method according to claim 44 , wherein the step of culturing in the presence of a ventralization signal transmitter is continued until a period when a rod photoreceptor precursor emerges.

46 . The method according to claim 35 , wherein the ventralization signal transmitter is a substance having an SHH signal transduction pathway promoting activity corresponding to 1 nM-10 μM SAG, or a substance having a BMP signal transduction pathway inhibitory activity corresponding to 0.1 nM-20 μM LDN193189.

47 . The method according to claim 46 , wherein the ventralization signal transmitter is SAG.

48 . The method according to claim 47 , wherein the concentration of SAG is 1 nM-10 μM.

49 . (canceled)

50 . The method according to claim 46 , wherein the ventralization signal transmitter is LDN193189.

51 . The method according to claim 50 , wherein the concentration of LDN193189 is 0.1 nM-20 μM.

52 . (canceled)

53 . The method according to claim 35 , wherein the method is performed in a medium free of 9-cisretinoic acid.

54 . A retinal tissue comprising a photoreceptor precursor rich in a rod photoreceptor precursor and/or a photoreceptor rich in a rod photoreceptor, wherein the retinal tissue is obtained by the method according to claim 35 .

55 . A retinal tissue comprising a photoreceptor precursor rich in a rod photoreceptor precursor and/or a photoreceptor rich in a rod photoreceptor, and a ganglion cell, wherein not less than 40%, preferably not less than 55%, of the number of the cells of the photoreceptor precursor and photoreceptor are rod photoreceptor precursors and rod photoreceptors.

56 . A retinal tissue that is able to mature into the retinal tissue according to claim 55 by culturing.

57 . The retinal tissue according to claim 54 or 55 , wherein not less than 50% of the layer structure of the retinal tissue forms a continuous epithelial structure.

58 . The retinal tissue according to claim 57 , wherein the retinal tissue has a diameter in the major axis direction of not less than 0.6 mm.

59 . A pharmaceutical composition for transplantation to a retinal tissue of a retina disease patient in need of transplantation, comprising a retinal tissue section cut out from the retinal tissue according to claim 54 or 55 .

60 . The pharmaceutical composition according to claim 59 , wherein the retinal tissue requiring transplantation is a region including the periphery of the macula and the outside thereof having a high proportion of rod photoreceptor precursor (Rod precursor) and/or rod photoreceptor.

61 . A method for treating a disease based on a disorder of a retinal cell or retinal tissue, comprising transplanting an effective amount of the retinal tissue according to claim 26 , 27 , 54 , or 55 , to a subject in need of transplantation.

62 . A method for evaluating toxicity or efficacy, comprising using the retinal tissue according to claim 26 , 27 , 54 , or 55 .

63 . A method for producing a completely matured retinal tissue that expresses S-opsin, L-opsin and/or M-opsin, comprising a step of culturing the retinal tissue according to claim 26 or 27 in a serum-free medium.

64 . (canceled)

65 . The method according to claim 63 , wherein the serum-free medium is a medium comprising a dorsalization signal transmitter.

66 . The method according to claim 65 , wherein the dorsalization signal transmitter is BMP.

67 . The method according to any one of claim 63 , wherein the serum-free medium further comprises a thyroid gland hormone signal transmitter.