DE1467915A1 - Process for the production of storable cell material - Google Patents

Description

DR. EYSENBACH

PATENT LAWYER PULLACH / MÖNCHEN

fag of inclusion t11. March Attorney's file number Κοβ-4-ρ

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description
to the

Patent application

"Technique for the production of storable cell material" Applicant Adan StASfZ, Morelos (Mexico)

The following priority is claimed: USA patent application Ser.Io. 351 220 rome March 11, 1964

The invention relates to a method for producing storable cell material with therapeutic efficacy duroh Preservation of cell tissue from mammals by means of cryogenic freezing or freeze drying Prepared preparations are in an individual specific Can be used in therapeutic treatments and also to increase milk production.

It is known that scientific research has long been concerned with the appendage organs of the uterus and the uterus

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busy and there are already TieXe works published been, dl · see the endoorine or hormonal functions and actives of such fetal attributes. For rubbing According to the present invention, the term "fetal attributes of the febrile process" refers to those attached organs the uterus of mammals, which "together old the tutus after penetration of the germ cell develop and sioh im generally separate from the uterus during the process of childbirth, regardless of whether these appendages are present at the end during pregnancy or artificially separated from the uterus. This cell material consists of fs.B. the end the egg shell, the chest water surrounding the fetus, the with the tutus tied yolk saok, the urine membrane, the cord, the cell material of the villi and the placenta and similar cells that surround the plaoenta and the tutus Membranes that form in the uterus during the development of the T5-tue and also the fluids that accompany the latter (e.g. the blood in the chest), you Baoh birth shedding from embryonic cell material of this type is well known} but it has already been pointed out that such embryonic cells are not just one contain significant levels of various hormones (e.g. estrogen, progesterone, gonadotropin), but essentially consist of single cells, which have a genetic structure similar to that of the mother organism and with the TU-tus ^ and the newborn individual that evolves from it genetically tolerable and at the same time with a high content of

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Antibodies en tereehen that the fetus against pathological Conditions of the Matter eu protect rermugen «The provision ron immunologically effective properties through the Embryonal et Ilen However, old cracks up on the ovarian gang, in which the bryonic cells (i.e. the embryonic cell tissue and the embryonic fluids) after the exit of the fetus from the uterus or even after a medical intervention that takes place Embryonic splitting (e.g. during a caesarean section) in the form of the xaoh birth be eliminated. In some men, however, Al Xmtaryonal cells of the afterbirth are not removed without being valued »there si · Tom dam are often eaten. The previous request Therefore, with regard to the properties of these embryonic cells and cellular tissues, Tor focused on theirs high content of different hormones and their role in Terlauf the Sohwangereohaft.

Xs has now been found to be derived from the fetal attributes of the existing embryonic pathway, regardless of whether it is after birth or during medical intervention Gate of natural expulsion of the fetus from the uterus contain, in addition to the high hormone levels found in practically all mammals, different active ingredients, which are not completely enlightened, but one specific to the mammalian indiTidues of the common genetic Origin (i.e. the mother from whom the embryonic cell material originates or the individual who develops from the fetus whose appendix is the embryonic cell material) have a directed effect and in this way indirectly

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They can be used therapeutically »B · was found in particular, let dl · Imbryomal« rush this kind, dl · la folgemdea al · «Ttabryo-Selbstabeonderuiif ·« · be without tmd di · sis * tob 4tr Inibryonalflüeeiglceit, dea fetus rail s, dtr cable cord "Vol d 'r placenta derived, Ib those IadlTldu" B _t di · ei oh au · dta enfthorigen f βταβ tntwi disgusting "and no oh naoh longer Intiriüklunfβ both no · iaaimolofiieh anepreoheade AlnrehrrealctloB Berrorruftn" 2a Gegemats mn of fact DAFL different, "iologleoh · Stuf etierextrakte β · 1 your Binrerleibung In a Indiriduellsn Organleaus aoraalerweiee" la · laaunolöflsoh aa- ■ preohende Aiwehrreaktlon sweep the einrerleibte herromifen biologleohs material are dl · Brabryo-Selbetabeonderungen _t "eetehend from dsn fetal attributes of a beetiamten Indlriduua's tone of this was not immediately repulsed eondsrn uagskshrt for surprisingly long times BurUokgefcalten and was auoh nooh then, when the "XndlTlduua" was concerned Entloklungsperlode after the VOtalstadiua has run through.

Accordingly, the aim of the invention is gruBdsätslloa la creating τοη biologieoh effective preparations for one indlTliual-epesiflsohs treatment tob S & alianoraanlemsn at all therapeutic indications

A particular aspect of the invention is the creation of a Preparation old indiridual-epeeifisoher »lrksaaksit bsi dsr treatment tob Tersohlsppten diseases (Xetastasls), Terleteung- • n, superficial and internal pathogenic conditions, DrOsea-

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malfunctions and the like.

Accordingly, the subject of misinformation exists for two purposes the rorigin geeohilderten and nooh others »below more detailed objectives in an »experienced for dividing up Clay storable cell material with therapeutic effectiveness by preserving clay cells from mammals by means of cryogenic freezing or freeze drying, which thereby It is well known that the starting material is embryonic Cells, partially from the egg, from the amniotic fluid, the yolk sac, the urine membrane, the cord, the villi skin, the placenta and / or the placenta blood is used.

Goal assignments are each originating from a single birth embryonic cells of the itttus appendages, in general the laoh birth can be taken separately.

The invention is based on the knowledge that this embryonic Cells nooh nooh longer periods of time according to the Sohwanger- ■ ohaft, from which they originate, are consistently effective if they can withstand destruction for such a long storage period have been made, although in the meantime both indicators have undergone considerable changes.

The embryonic cells mentioned above can be used for this purpose through a defrosting process (e.g. by freezing at temperature) or treat by the usual freeze drying. Man

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The embryonic cells passivated in this way can suspend considerable loss of their individual, epesifical therapeutic efficacy for a long time in a manner that is similar to the storage of semen material for subsequent nutating in artificial insemination, as already indicated It is most surprising that the embryo self-secretions, that is, the fetal envelopes, ietus fluids and plaoenta cells _; retain their ability to »not trigger an immunologically responsive defense reaction in those mammalian individuals for whom these embryo cells are specific. Although all the indications suggest that under certain circumstances the embryonic cells are "suitable" for the treatment of the feed organism from which they originate, it should be pointed out that these cells provide their main support in the treatment of the child organism resulting from this pregnancy The biological incorporation of these embryonic cells can be carried out in the form of a coherent cell tissue (e.g. from the cord, the egg envelope, the villus membrane or the placenta) as an implant, either on its own or in combination with an additional implant made of nonembryonic cell tissue, for example as a reduction of injuries in the treatment of burns, superficial skin injuries, ulcers or the like, or only as implantations in nooh v present organs

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! «• ok · exercise tint · invigorating effect on 41 · lymphatic and aruse-controlled defenses.

for the latter two, β is also possible, di-embryonic Cells by oral ingestion or by injection. The- ·· cells are then suspended in a liquid carrier be, in Terträgliohen pharmakologisohen questioning materials for di · oral · administration or also in fixed tone can be used.

Ie has shown that the preparations produced according to the invention old especially good effectiveness for rereohledene therapeutieohe Can be used for two purposes if the embryonic cell material is not input directly to the individual tee should be treated, but da8 ee instead of a • Large mammals are taught to tone the secondary cells Darreiohung be produced for the mammal to be treated * Ie has shown that this technique: especially with the Treatment tone metastasis is useful.

In this case the metaetatieohe becomes in a first state Mammal treated in such a way that primary cells are introduced into a second mammal that are from one mammal indiriduum originate with the first mammal in the weeentliohen possesses the same genetic structure, namely either the first mammal itself or, Tore eye-wise, its cutter ι at the same time or adjacent to the inner abdomen of the embryo

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Basal primary cells become the second mammal auoh metastatlaohe Cells of the first mammal · incorporated. You τοπ dem Secondary cells generated by a third mammal are in this way taken in the form of a serum extract from the blood, the lymphatic fluids and / or the milk and entered the first mammal.

It has proven to be particularly advantageous that those of the indiriduum to be treated or those mentioned above The embryonic embryos incorporated into the mammal and treated according to the invention are capable of producing milk in this embryo to demand or bring in Oang. More precisely, 1st to establish that the incorporation of embryonic cell material, e.g. by implanting cord or placenta cell tissue into the uterus of a female mammal whose Milk production promotes one by increasing milk production already lactating or by causing lactation in an uncovered her. If at the same time a metastatic Additional implantation is carried out at the second her, it happens that the milk in the treatment of the metastatic State of the individual from whom the cancer cells originate, has a therapeutically beneficial effect. In general, they are obtained, for example, from the body fluid of the second animal secondary cells originating from the property, at least for • an initial period, no immunologically responsive defense reaction evoke, although the embryonic cell material belonging to this individual, which is involved in its formation

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leading Sohwangersohaft has developed "bti dtr treatment of this" individual is not directly used, la • I have further shown that the preserved bmbryo-self-secretions obtained according to the invention, ie for example the cover, the early water, the yolk sac, the urine, the cord, the villi skin (base, villi and deposition), the PIa-ο ent a and the plaoenta blood, can be used as appendage attributes of the fetus without eliciting an immunologically responsive defense reaction for the therapeutic purposes mentioned above. This cell maivial consists essentially of the same genetic structure as that of the cutter and the child and therefore contains a very high defense potential (Hofbauer cells, etc.). The basic idea of the present invention consists in AaS of this embryonic cell material, which has previously been lost, is now preserved as soon as it is removed during the birthing process or during a medically performed birth. The Matrial is then in a by freezing or by Gefrietrooknung reached passive state, if necessary for years, lifted up to the time at which this cell material for the above described implantation purposes or -F in ^{^} ovm of homogenised Injektionspräpsraten - as Agent for the treatment of lymphatic or gland-controlled malfunctions is used. Thus, these extracts can be used, for example, in the treatment of the sohädliohen progressive reduction of the lymphoid cells, both in the developing organism (ie in the child) and in the mother organism when there are symptoms of the

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Aging or in various pathological conditions will. Al-We ind the durable ones produced according to the invention Embryo self-secretions are said to be diseases and / or malfunction of various kinds, e.g. in Oollagen diseases, metastatic processes in tuberculosis or cancer, dysfunction Tone hematopoietic cells (e.g. in leukemia) or Injuries caused by any of the above diseases or by mechanical or other pathological conditions will. Pas embryonic cell material can be compared to normal Birth through the recovery of the laughing birth, the fotus envelopes, the fruoht bladder fluid kex? and also of the placenta blood during deflation of the fetus} but it can also be obtained from the fetus Implementation of the Kaisersohnittes can be won. The embryonic cells can be axed in ritro or they can also without further development near their passivation by freezing or freeze drying can be used in the aforementioned manner. It may be that some of the above The therapeutic successes mentioned are partly due to the presence of therapeutically active hormones in the embryonic cells will; However, it can be assumed that the individual-specific Efficacy provides evidence that the primary therapeutic effect is in the genetic combination is due to the various cells and cell products produced during pregnancy, if also added must be that the effect mechanism is by no means clarified One might suspect, however, that bmbryo self-secretions various elements of the reticuloendefelial system in addition to various hormonally effective components (e.g. ginatropin

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ami 4tr villi skin, estrogen, progesterone) and aoetyloholin, which enable the embryonic cells to be absorbed without the body defending themselves. Accordingly, it is an important feature of the present invention that the cell material consisting of the xmbryo self-secretions, such as the enveloping skin etc., is collected separately for each indiriduum at birth, made durable and stored in order to be able to use it at a later point in time, which may possibly arise Years after the Q- birth can be eligible for the treatment of this indirect EU.

You mentioned goals, features and objectives of the invention The following examples of the principles used in the preparations made according to the invention are intended to result in gate parts can be used for various therapeutic purposes, are explained in more detail.

I. Stimulation of lactation

IiAi 3 year old Jersey cow that is near a normal lactation Miloh was given an intramuscular injection of ron Plaoenta lines in homogeneous solution, with the Plaoenta lines originated from Plaoenta material that was littered by the same cow after the calving which was about to begin when pregnant had been. The Plaoenta cells were immediately after throwing been frozen and slowly returned to room temperature before injection brought and homogenized duroh a Yermengung process

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been. The success consisted in an increased relief for 15 positions, in which the increase in milk output was sat above the average milk output during this 15 position * On the first day after the injection of the placenta ropes, an increase in milk output of 2 liters was observed and this increase in Miloh output sank after 8 days to a daily amount of clay 0.5 liters. It was injected into the animal's soot and 5 com of the homogenized PIaoenta cell material was used. Three months later, 3 com of freeze-dried placenta cells derived from the above-mentioned placenta were injected into the swan. An increase in milk yield of around 800 to 1000 com per day was achieved. It was found that the freezing process can be most effective if the plaoenta cells are washed / separated under sterile conditions and only subjected to a freezing process, but the low temperature freezing process is carried out at significantly lower temperatures. The Sonell freezing process was carried out in such a way that the temperature Eunäohet was lowered rapidly to about 5 ⁰ O and then to -5 ⁰ O, while the subsequent low- ^{temperature freezing was carried out slowly to a temperature Ton -40 0} O.

II. Induction of lactation in undoed animals

A 3 year old undetected sheep (Merino strain) was in his Uterine finds with implantations τοη human placenta and

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■ abtlsohnur-Zsllgtwebe (j «welle 1,3 den) Tersehen. Dae iaplanted Zellgew ·! »· Became during a period of time τοη 6 Woohen maintained, and it showed that it was under great conditions the implantation region τοη dta animal TOllet Permanently assimilated had been. During the period τοη 6 woohen tine-induced lactation with a milk production τοη approximately 5 oom per day.

III Treatment of a metastatic disease

line 27-year-old trust of the white Sasse old the characteristics of obesity _t who had uterine cancer with endometriosis »was treated with secondary cells» which had been accustomed to implantation τοη metastatisohtm cell tissue of this individual in the uterine wall of the sheep described under II. *. The implantation of the metastatic material formed an additional implant ("eupergraft") that was implanted in the sheep together with the Habelsohnur and placenta cell tissue. The cell tissue of the placenta and the cable cord were removed from a pregnancy of the metastatic patient and had been stored for a period of τοη 3 months under the conditions of the freezing method »which is described under I ·. An implantation τοη attastatisohta material was also introduced into the tail, and 3,000 dta tissue from tissue cells, which were suspended in 30% of an inert liquid, were applied intra-abdominally. Both from dta arterial and renal blood dts

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It also consists of lymphatic fluids and fluids A serum was produced for his Miloh. The implantation of the cancer cells acted as an increase in coloetral production of the sheep by about 30,000. The serum became the patient duroh intramuscular injection in 2 oom dose three times in one Where oh β taught. In addition there were 20 oom me of the sheep given orally three times a week. The patient showed a general improvement in her health with a reduction in the Obesity, during. The metastatic area in the course of the Treatment showed a considerable decrease in its extent.

IT · Treatment of relocations

I am a 3 year old merino sheep that emerged from underwater The Bauohdeoke suffered »was provided with a cellular tissue closure made of meneohliohem labelohnurmaterial, which was separated lengthwise and had been spread out to spank the access opening. Already after 6 hours a remarkable one appeared Healing of the injury and the implanted · Eautstüok was maintained for a period of 6 weeks with subsequent assimilation. Bs also showed that the injury Anatomically properly healed and, with regard to the strength of the healed area, compared to the usual baht spuds was superior. The Habelsohn's material used in this medical treatment was among those for 3 months Conditions of the deep freezing method described under I.

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been stored.

T. wound treatment

2 lin male IaIb was in an area tone 10 om with a

Skin reliefs that are both dermal and subcutaneous fabric length of one side of the jaw. on Cell tissue of the villi and the outer layer of the skin with some urinary material was applied to the wound area. Inside clay During β days there was a great clearing, while a similar one sores symmetrically on the other side of the veal sausage had been attached »which took t weeks to heal. At the wound site treated with embryonic cell material There was already a visible sore sole plunging after C hours an "wobu" needed 24 hours for the untreated wound became. Moreover, see that the latter was attacked by maggots »whereas the treated sore clay remained free of this infection» suffered no irritation and an anatomically perfect healing region with significantly lower generation τοη compared to the KU of the untreated area Barbel tissue formed. The cell material used was obtained from the membranes surrounding the fOTus, which were treated with the XaIb at his birth had been ejected 3 months surer. The cell tissue τοη the placenta was removed, washed under sterile conditions, τοη muscle fibers cleaned, and frozen then subjected to the animal freeze process. A second fur of the cell tissue was freeze-dried, stored for 3 months and then inserted along the xanten of the implanted material

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been brought.

. Curative treatment of an ulcer

A 34 year old white woman with good general Health condition suffered from a superficial ulcer with Blood vein widening ι eie was treated with Plaoentaaellen, dl · Sound of a woman who was just pregnant. The cells had been stored after freezing for more than a year The placentas were applied to the surface every 2 weeks and complete healing was observed after 6 weeks. Ie also became a general exertion of the physiological Characteristics of the patient and also a psychological effect in the form of increased happiness.

Due to the above-described effects of the invention The cell material produced is to be assumed that the Taoh people have a much larger area of application the re-settled areas will be found.

Dr.lyiHa

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Claims (9)

Day of the linrelohungi 11th March 1965 lawyer "afc *" net worth it! HoflM »? U67915 1. Terfahren * ur production of storable cell material with therapeutic effectiveness by preserving τοη cells also for mammals by means of low-temperature cooling or freeze-drying, characterized in that embryonic poultry attachment cells, roreugeweiee, are used as the outgrowth material au · der lihülle, dtm fruohtwasstr, dta Dottereaok, the Urnhaut, dtr Kabeltohnur, dar fottenhaut, the Plaoenta and • sw. odtr dt · Plaoenta blood rerwendet. 2. retracts, NaOH Anipruoh _y 1 characterized gekennaeichntt that one · al Auafangamaterial jeweile au · a eineigen Oeburt etaaaende Fetal appendages separated from each other. 3 * Proceeding according to one of the tests 1 or S ₁ without knowing that the preservation takes place by means of low-temperature freezing. 4 * Driving according to Anepruoh 3, daduroh gekennselohnet , dal dl · Durability medium · Low temperature freezing In 2 ■ tat · * «rf« Igt,? »·· 1 dl · * ret · Level« Uw Aikttmluag Hl « ₉ tm -5 * 0 eohnell and dl · further aluminum cooling to about -40 * 0 takes place. 90981 1/1162 φ 1% H67915 5. Driving near a dtr Aneyrüohe 1 and 2, thereby g · ktoniiiohntt that the preservation by elm «a ■ Well-known freeze drying procedure is carried out. 6. retracts, NaOH any preceding Aneprüehe "daduroh gekennüelohnet _f defl ale Auegangeaaterial ββ intact embryonic tissue is used · 7. Terfahren naoh one of the Torhergoing claims, thereby marked that the cell material made durable Gate of its application in a liquid vehicle dieperglert. 8 · ferfahrea "for irhöhung lactation, daduroh geke η η ■ elohnet, DAJ a female mammal which was the Sohwangereohaft a Säugetieree ml · Retus attachment organs entwlekelten embryonic cells" Torsugewelee from the lihülle, the fruchtwas8er _f the Dottereaok which Harnhaut, the Vabelsohnur, the villi skin, the placenta and baw _a or dee Plaoentaelutee are incorporated, 9 · Terfahren according to claim 8 »daduroh marked, d «0 the cell material used before its application one of the usual Terfahrea, in particular about fleftemperatareinfrlerung oitr 8efriertroeknung in paintable ^ storable is brought into shape. 909811/1162 BAD -S- 10 · Process for the preparation of a serum for therapeutic Application toeifstatisohem disease state, thereby characterized in that an embryonic cell material produced according to one of claims 1 Ils 7, that comes from a sick human individual or its mother, a mammal capable of producing milk implanted, this mammal also metaatatisohes cell material of the diseased human individual and which developed the clay in the mammal after this treatment Secondary cells processed into a serum in a manner known per se, that can be supplied to the sick human individual. Dr EytHa 9 0 9 8 11/116 2