Efficient production ofoffspring in wild-derivedstrains of mice (Biol Reprod inpress)
Birth of offspring from ectopicallytransplanted PGCs (Biol Reprod2011)
ICSI with sperm derived fromcultured testes (Nature 2011)
Establishment of rabbit iPS cells(J Biol Chem 2010)
Establishment of ES cells fromimmature oocytes (Stem Cells2011)
Improvements of efficiency insomatic cell nuclear transferby correcting aberrant Xistexpression in cloned embryos(Science 2010; PNAS 2011)
(Presented by Keiji Mochida)
Implantation
0
20
40
60
80
100
ICSI ELSI ROSI
Overall
0
5
10
15
20
25
ICSI ELSI ROSI
Survival
0
20
40
60
80
100
ICSI ELSI ROSI
Cleavage
0
20
40
60
80
100
ICSI ELSI ROSI
Birth
0
10
20
30
40
50
60
ICSI ELSI ROSI
Birth Overall efficiency
Survival after injection Cleavage Implantation
Ogonuki et al., PLoS One 2010
B6
ELSI: elongated spermatid injection; ROSI: round spermatid injection
・
・
・
Ogonuki et al. PLoS One 2010
・
・
・
Overall (Offspring/oocytes injected)
0
5
10
15
20
25
ELSI ROSI
(%)
Sperm Elongated Roundspermatid spermatid
・
・
・
Ogonuki et al. PLoS One 2010
・
・
・
Overall (Offspring/oocytes injected)
0
5
10
15
20
25
ELSI ROSI
(%)
Sperm Elongated Roundspermatid spermatid
Genetic conservation Poor reproductive performance by aging
Motility failure of frozen sperm due to cryoinjury
Simple cryopreservation of male gametes
Fertilization of cumulus-free cryopreserved oocytes
Gene function analysis Sperm-oocyte fusion protein KO mice (CD9, Izumo)
Cumulus-deficient KO mice
Spermatogonial stem cell transplantation Allogeneic spermatogonial transplantation
Confirmation of clonal origin of spermatogonial colonies and many…
Gene therapy Rescue of Sl/Sld mice by virus-mediated gene therapy
Production of interspecies F1 C57BL/6 x Spretus
Transgenesis Sperm-mediated transgenesis by ICSI
Speed congenic Use of first wave spermatogenic cells
Analysis of sperm factor Sperm-borne oocyte-activating factor
In vitro manipulation of immature germ cells ES cell-derived sperm
Spermatids derived from in vitro-meiosis
PGC ectopic transplantation
B6;129S-Gtrosa26LghDDXC-PSMXA-1
C3.KOR-Apoeshl
CXDD-MSTS(tremor)NOD.R209-LineTNOD-Spretus Congenic 18-1C57BL/6J-TgN(b-actin-hBACE1)riken2Floxed Sox2 mice
mel18-/-CNR303cp/v11ve1KAFUBICard15KO129 x B6-G129 x B6-GIVCS.129P(B6)-Akr1c18<tm1Mni>SMXA-45SMXA-33SMXA-34NZB.Cg-Cd180<tm1Kmiy>B6;CB-Rab27a<Gt(pU-21T)58Imeg>/RbrcFVB129P2(B6)-Mmp2<tm1Ito>B6C3-Tmem50b<Gt(venusTrap_fJAV0017-B)B6.Cg-Tg(Scx-GFP)1StzrC57BL/6-Tg(CAG-Nup85)3-3SushSperm with a bent neck
from a TG mouse
Genetic conservation Poor reproductive performance by aging
Motility failure of frozen sperm due to cryoinjury
Simple cryopreservation of male gametes
Fertilization of cumulus-free cryopreserved oocytes
Gene function analysis Sperm-oocyte fusion protein KO mice (CD9, Izumo)
Cumulus-deficient KO mice
Spermatogonial stem cell transplantation Allogeneic spermatogonial transplantation
Confirmation of clonal origin of spermatogonial colonies and many…
Gene therapy Rescue of Sl/Sld mice by virus-mediated gene therapy
Production of interspecies F1 C57BL/6 x Spretus
Transgenesis Sperm-mediated transgenesis by ICSI
Speed congenic Use of first wave spermatogenic cells
Analysis of sperm factor Sperm-borne oocyte-activating factor
In vitro manipulation of immature germ cells ES cell-derived sperm
Spermatids derived from in vitro-meiosis
PGC ectopic transplantation
medium
nNo.
pregnant
Sperm GL-PBS 4 4 50 34 (68.0) 11 (22.0)
NIM 2 2 20 19 (95.0) 13 (65.0)
El. Spermatid GL-PBS 1 1 10 3 (30.0) 1 (10.0)
NIM 4 3 39 12 (30.8) 7 (17.9)
Cells used
RecipientNo. transferred
(Total)
No. (%)
implant
No.(%)
offspring
Ogonuki et al., PNAS 103: 13098-13103, 2006 in collaboration with Dr. Martin Fray
o
(provided by Dr. Iwaki, Tokyo Jikei Medical University)
Feb. 13, 2006
Aug. 28, 2006
Ogonuki et al., PNAS 103: 13098-13103, 2006
No. used No. pregnant
BALB/c-nude 106 81 (76.4) 81 6 4 42 (51.9) 17 (21.0)
C3H/He 108 97 (89.8) 97 7 4 34 (35.1) 12 (12.4)
Strain of maleNo. (%)
offspring
Recipient femalesNo. embryos
transferredNo. (%) implant
No. oocytes
that survived
inejction
No. (%) oocytes
cleaved
Mar. 2, 2006
Genetic conservation Poor reproductive performance by aging
Motility failure of frozen sperm due to cryoinjury
Simple cryopreservation of male gametes
Fertilization of cumulus-free cryopreserved oocytes
Gene function analysis Sperm-oocyte fusion protein KO mice (CD9, Izumo)
Cumulus-deficient KO mice
Spermatogonial stem cell transplantation Allogeneic spermatogonial transplantation
Confirmation of clonal origin of spermatogonial colonies and many…
Gene therapy Rescue of Sl/Sld mice by virus-mediated gene therapy
Production of interspecies F1 C57BL/6 x Spretus
Transgenesis Sperm-mediated transgenesis by ICSI
Speed congenic Use of first wave spermatogenic cells
Analysis of sperm factor Sperm-borne oocyte-activating factor
In vitro manipulation of immature germ cells ES cell-derived sperm
Spermatids derived from in vitro-meiosis
PGC ectopic transplantation
First wave round spermatidsin a 17 day-old testis
Miki et al., JRD 2004
A pup born from a 17 day-oldspermatid
Genomic DNA cDNA
1 2 3 4 5 6 7 8M 9 10
SpermatidInjection
H19
JF1B6
H19, Igf2, Meg3, Igf2rIVF
(B6 x JF1)
01020
3040506070
8090
100
0 20 40 60 80 100 120
Markers
homozygousforB6(%)
10
20
30
40
50
60
70
80
90
100
0 50 100 150 200
Markers
homozygousforB6(%)
80
85
90
95
100
0 50 100 150 200 250 300 350
Markers
homozygousforB6(%)
Gdf5
EdnraEGFP
Kitlg
RELMb
0
20
40
60
80
100
0 50 100 150 200 250 300
Markers
homozygousforNOD(%) (70/72)
(63/72)(60/72)
(38/72)
(0/72)(0/72)143d
20d
45d 43d44d
(72/86) (72/86)
(85/86)
(77/86)
(86/86)41d
43d
162d97d20d
43d
43d
(0/86)
(56/86)
(79/86) (86/86)
(38/176)
(123/176)
(154/176) (163/176)(176/176)
44d
42d41d
43d
Ogonuki et al., PLoS One 2009
Genetic conservation Poor reproductive performance by aging
Motility failure of frozen sperm due to cryoinjury
Simple cryopreservation of male gametes
Fertilization of cumulus-free cryopreserved oocytes
Gene function analysis Sperm-oocyte fusion protein KO mice (CD9, Izumo)
Cumulus-deficient KO mice
Spermatogonial stem cell transplantation Allogeneic spermatogonial transplantation
Confirmation of clonal origin of spermatogonial colonies
Gene therapy Rescue of Sl/Sld mice by virus-mediated gene therapy
Production of interspecies F1 C57BL/6 x Spretus
Transgenesis Sperm-mediated transgenesis by ICSI
Speed congenic Use of first wave spermatogenic cells
Analysis of sperm factor Sperm-borne oocyte-activating factor
In vitro manipulation of immature germ cells ES cell-derived sperm
Spermatids derived from in vitro-meiosis
Ectopic PGC transplantation